Tag: gDNA

DOI: 10.18129/B9.bioc.gDNAx   Diagnostics for assessing genomic DNA contamination in RNA-seq data Bioconductor version: Release (3.18) Provides diagnostics for assessing genomic DNA contamination in RNA-seq data, as well as plots representing these diagnostics. Moreover, the package can be used to get an insight into the strand library protocol used and,…

Genetic characterization For risk stratification and identification of genetic aberrations suitable as MRD targets, diagnostic material from 133 children with primary AML was examined (see “Methods” and Supplementary Table 3). The presence of selected fusion genes and mutations was prospectively investigated using (q)RT-PCR and sequencing. A primary genetic aberration was found…

Plasmid construction All the constructs used in this study were generated with standard cloning strategies, including PCR, overlapping PCR, oligo annealing, digestion and ligation. Primers were purchased from Genewiz. The plasmid sequence was verified by Sanger sequencing. The pCAG-spike(D614G)-GFP11-mCherry plasmid was modified from Addgene plasmid 158761. Briefly, GFP11 and mCherry…

Summary Despite whole-genome sequencing (WGS), many cases of single-gene disorders remain unsolved, impeding diagnosis and preventative care for people whose disease-causing variants escape detection. Since early WGS data analytic steps prioritize protein-coding sequences, to simultaneously prioritize variants in non-coding regions rich in transcribed and critical regulatory sequences, we developed GROFFFY,…

Working with NCBI downloadable Datasets 0 Hi all, I’m an postgraduate student currently working on an assigment in the field of “Analysis Molecular Data”. We’ve been instructed to examine polymoprhism in the promoter of the gene MMP3 in humans, and how that might affect expression and causality for genetic disorders….

The turnaround time for HLA and genome sequencing varies based on the sequencing type, sample quality, and number of samples. We typically estimate turnaround between 10-15 business days for smaller projects, from the time your samples arrive in the lab to when we deliver results. Important Notes DNA extraction is…

A viral RNA produced during Φ80α-vir infection activates Ssc-CdnE03 in vitro. a, Detection of phage propagation after spotting tenfold dilutions of the lytic DNA phages Φ80α-vir, ΦNM1γ6, ΦNM4γ4 and Φ12γ3 onto lawns of S. aureus RN4220 harbouring a plasmid expressing an incomplete (Ssc-CdnE03 alone) or intact Ssc-CBASS operon. b, Thin-layer…

I’m running into a error that I haven’t managed to debug so far. The job runs for a few seconds, and then stops writing files. The errors are not always exactly the same, nor they are happening at the same point of the subsetting process. Here is a representative example,…

Genome editing technology stands at the forefront of current life science research. However, for its successful translation into clinical applications, the accurate detection of off-target effects is indispensable. Assessing and mitigating off-target effects correctly is a pressing concern. The issue of off-target effects in CRISPR-Cas9 gene editing technology has been…

Barcoding of native genomic DNA libraries We advise all customers to read and consider the information in the Kit 14 sequencing and duplex basecalling info sheet before starting this protocol Requires the Native Barcoding Kit 96 V14 (SQK-NBD114.96) PCR-free protocol Using up to 96 barcodes Allows analysis of native DNA…

Introduction Since the emergence of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) in 2019, this coronavirus has spread to more than 200 countries, cumulatively infecting 672 million people and causing 6.84 million deaths. Moreover, the emergence of numerous variant strains (α, β, γ, δ, and Omicron) presents a serious challenge for…

Animals for single-cell sequencing Adult samples of the aerial web-building spider (Hylyphantes graminicola) were collected from Anci district, Langfang, Hebei, China (39° 31.90’ N, 116° 38.15’ E) between September and October 2020. Collected spiders used for brain dissection were housed individually in a glass tube (Φ12 mm × 80 mm) at temperature- and humidity-controlled condition (24–26 °C and 50–60%…

doi: 10.1016/j.lrr.2023.100393. eCollection 2023. Affiliations Expand Affiliations 1 NSW Health Pathology, Liverpool Hospital, Liverpool, NSW 2170, Australia. 2 Ingham Institute for Applied Medical Research, 1 Campbell St, Liverpool, NSW 2170, Australia. 3 Centre for Circulating Tumour Cell Diagnostics & Research (CCDR) at the Ingham Institute for Applied Medical Research, Liverpool,…

Introduction Cystic fibrosis (CF) is a lethal, autosomal recessive disorder caused by mutations in the CFTR gene that results in aberrant transport of chloride, bicarbonate, and sodium ions (Kerem et al, 1989; Riordan et al, 1989; Rommens et al, 1989; Tabcharani et al, 1991; Poulsen et al, 1994; Stutts et…

BAC plasmids Human (CH17-203N23, CH17-449P15 and CH17-339H2) and mouse (RP23-51O13, RP23-75P20 and RP23-204E8) BACs were purchased from BACPAC Resources Center. Yeast–bacterium shuttle vector pLM1050 was modified by L. Mitchell based on a previous study28. pWZ699 was constructed by inserting a cassette containing pPGK-ΔTK-SV40pA transcription unit and the Actb gene into…

A large deletion mutation of 2811 bp of gDNA was introduced into zebrafish vtg2 via CRISPR/Cas9 genome editing. A schematic representation of the general strategy for CRISPR target design and application is given in Fig. 1A–C. The introduced deletion involved 1692 bp of the vtg2 transcript, encoding 564 aa of its respective protein, and…

RT-qPCR primer specificity can be measured using a bioinformatics workflow that involves analyzing potential mismatches, cross-matches, co-amplification of multiple gene splice variants, and sub-optimal amplicon sizes in silico. This workflow utilizes publicly available resources such as NCBI Primer BLAST, in silico PCR in UCSC genome browser, and Ensembl DNA database…

PCR can be performed on embryo spent culture medium to detect genetic information. Studies have shown that miRNA, such as miR-191-3p, can be detected in the culture media of human embryos and may serve as a molecular marker for assessing embryo competence in a non-invasive way. Additionally, the ratio of…

Instruments MISEQ ILLUMINA NGS sequencing platform that allows to generate up to 25 million reads from 25 to 300 nucleotides in paired end mode (PE 2X300)  MinION ONT Portable real time device for DNA and RNA sequencing by nanopore technology 3730xl GENETIC ANALYZER Capillary electrophoresis platform for high throughput genetic…

Your browser does not support the video tag. Terabases of data 1 – 48 flow cells Flexible, on-demand sequencing High-performance compute Analyse data in real time with 4 NVIDIA A-Series GPUs Up to 13.3 Tb data* * (Theoretical max output when system is run for 72 hours at 400 bases…

Introduction Antimicrobial resistance is a hidden global pandemic that shattered over 4.9 million people in 2019 alone, and the burden is highest, mainly in low-resource settings.1 Drug-resistant tuberculosis (DR-TB) caused by Mycobacterium tuberculosis (Mtb) complex (MTBC), which is resistant to one or more anti-TB drugs, is a leading global public…

The veil on genetic information was lifted by the advent of gene cloning and sequencing technologies in the 1970s. The discovery and purification of bacterial nucleases capable of cutting DNA at specific sequences and ligases capable of joining them enabled DNA from any species – or mRNAs converted to complementary…

Genomic DNA or gDNA is chromosomal DNA, in contrast to extra-chromosomal DNA such as plasmid DNA. Genomic DNA contains all the biological information to be passed on to the next generation. Most organisms have the same genomic DNA in all their cells, however only certain genes are active in each cell. Genomic DNA…

Control assays and synthetic DNA templates were designed to facilitate the assessment of the key experimental factors impacting your real-time PCR results. DNA Contamination Control AssayUse the PrimePCR DNA contamination control assay to determine if genomic DNA (gDNA) is present in a sample at a level that may affect PCR…

The FML-seq method The protocol for FML-seq comprises only three steps (Fig 1A). First, genomic DNA (gDNA) is digested by a methylation-dependent restriction endonuclease that cuts at a certain distance from the 5-methylcytosine or 5-hydroxymethylcytosine in its motif and leaves a short overhang (10). Second, a master mix is added…

 As Marvel superheroes traverse the multiverse to save the day, genomics researchers are our superheroes as they navigate the daunting multiverse that is biology. The complex and dynamic interactions between the genome sequence, its epigenetic regulation, and their combined effects on transcript expression and splicing are fundamental to our understanding…

Introduction: Natural killer (NK) cells are cytotoxic innate lymphocytes that eradicate tumor cells. The induction of durable anti-tumor immune responses by NK cells is a priority for cancer immunotherapy. Although cytosolic DNA sensing plays a crucial role in initiating anti-tumor immunity, the role of NK cell-intrinsic STING signaling is unclear….

1. Introduction Newborn screening (NBS) plays a vital role in healthcare systems for the prompt identification of individuals who may develop one of a set of rare, but severe health conditions [1]. In the UK, for example, dried blood spot (DBS) specimens are routinely collected at 4–5 days of age…

In a proof-of-concept study published in The Lancet Microbe, researchers from Australia developed a point-of-care assay for the rapid detection of mpox virus (MPXV) in clinical samples based on isothermal amplification along with clustered regularly interspaced short palindromic repeats (CRISPR) CRISPR-associated protein (Cas) technology. The assay was found to be…

A proof-of-concept study published in The Lancet Microbe introduces a promising point-of-care assay for the rapid detection of monkeypox virus (MPXV). The researchers from Australia developed the assay based on isothermal amplification and CRISPR-Cas technology. With its high sensitivity and specificity, the assay performed well when compared to the gold-standard…

All the extractions, dilutions and library preparation for sequencing were carried out inside the clean room environment of ISO (International Organization for Standardization) level 5 (ECSS-Q-ST-70-55C standard)32. An ISO 5 Class clean room is a semi-closed ultra clean environment that utilises High Efficiently Particle Air (HEPA) filtration systems to maintain…

PacBio HiFi long-read sequencing is one of the most advanced sequencing technologies for generating groundbreaking discoveries that can expand our understanding of human health. In case you missed it, we recently published a round-up of some of what we think are the most significant new research publications to employ PacBio…

Stem cell and cerebral organoid culture conditions Feeder-free hES cells or iPS cells were cultured on hES cell-qualified Matrigel (Corning, catalogue no. 354277)-coated plates with Essential8 stem cell medium supplemented with bovine serum albumin (BSA). H9 embryonic stem cells were obtained from WiCell. Cells were maintained in a 5% CO2 incubator at 37 °C….

Human plasma collection Fifty patients who were assigned for CPB surgery between May 2021 and February 2022 at the Department of Cardiac Surgery, Southern Medical University Nanfang Hospital (Guangzhou, China) were recruited given their increased risk of intestinal I/R injury. In patients undergoing cardiac surgery, CPB is potentially responsible for…

Strain identification The strain Phoma sp. CNCM I-4278 was previously isolated from the rhizosphere of Nicotiana tabacum (cv Xanthi, Solanaceae) grown under controlled conditions17. The fungus was identified according to the closest similarity of its 18 S rRNA sequence (HM16174323) with those present in GenBank by that time and it was…

The research in this study was conducted under the ethical approval of the Danish Regulatory Authority under project license 2018-15-0201-01520. Cell culture and differentiation assays WT, MCM2-2A, MCM2-R and POLE4-KO mouse ESCs used in this study were derived from the male, E14JU cell line with a 129/Ola background81. Genome editing…

Animals The gene targeting experiment was conducted on cynomolgus monkeys, which served as the experimental subjects. These monkeys were housed at Guangdong LANDAU Biotechnology Co. Ltd and TOPGENE Biotechnology Co. Ltd, where they received unrestricted access to water and were provided with a standard diet in accordance with standard care…

Sleeping Beauty transgene overexpression Bidirectional Sleeping Beauty plasmids encoding the codon-optimized cDNA of the gene of interest (FLT3, KIT, CD123) or their mutated variants were cloned downstream of an EEF1A1 promoter. The same construct included a reporter cassette comprising a fluorescent protein (mCherry or mTagBFP2 for FLT3 and KIT, respectively)…

Skaletsky, H. et al. The male-specific region of the human Y chromosome is a mosaic of discrete sequence classes. Nature 423, 825–837 (2003). Article  ADS  CAS  PubMed  Google Scholar  Porubsky, D. et al. Recurrent inversion polymorphisms in humans associate with genetic instability and genomic disorders. Cell 185, 1986–2005 (2022). Article …

1. Introduction Crops such as rapeseed, along with vegetables like cabbage and Chinese cabbage, all of which belong to the Brassica genus, are widely cultivated across the world. Chinese cabbage, in particular, holds significant commercial importance as a vegetable extensively consumed in Asian countries [1,2,3]. Chinese cabbage thrives well in…

In a recent article published in BMC Medical Genomics, researchers recruited a cohort of 25 Chinese genodermatoses patients to examine the genetic etiology by whole-exome sequencing (WES). They evaluated the clinical utility of WES as a tool with the potential to expedite the discovery of the genetic variations underlying all skin…

Development of safe and effective viral specific gRNAs CRISPR gRNA designer tools were used to identify gRNAs that target LTR and Gag with minimal chance of unintended effects for the previously described HIV-directed guides [6, 10]. Previous research demonstrated efficacy in targeting HIV in culture and using an infected humanized…

Pan-cancer WGS data sources GrCh37/hg19 BAM alignments for 2,489 primary tumour and matched normal whole-genome sequencing data were obtained as previously described18. In brief, 989 tumour–normal (T/N) pairs were obtained from The Cancer Genome Atlas (TCGA) Research Network (Genomic Data Commons at portal.gdc.cancer.gov/, accession: phs000178.v11.p8). Additional WGS data were obtained for 874 T/N pairs…

Highly-accurate HiFi reads produced by PacBio long-read sequencers –such as the Sequel IIe and Revio systems– enable researchers to access entirely new levels of contiguity, completeness, and accuracy when assembling genomes. The sequencing capabilities of these long-read platforms are particularly beneficial for microbial genome assembly. The superiority of PacBio assemblies…

Sample selection Samples from two mass gravesites located near one another and dating back to the Second World War were selected because of their challenging yet relevant characteristics in forensic genetic examinations (skeletonized human remains, long postmortem interval, and DNA degradation). The samples were the diaphyses of femurs, cut below…

Novogene’s Revio Data: Achieving Maximum 110Gb HiFi Reads per Cell! Novogene, a leading provider of genomic sequencing services, has recently installed the latest PacBio Revio long-read sequencing system in its US and Chinese laboratories. This cutting-edge technology offers throughput of 360 Gb of HiFi reads per day, a 15× increase…

Morphological observation of continuous in vitro cultured B. gibsoni (Wuhan isolate) Babesia gibsoni was successfully cultured in vitro in 20% serum. After splitting, parasitemia reached 10% ± 1.5% on day 3 (Fig. 1A). Fig. 1 Changes in parasitemia and morphology of in vitro cultured B. gibsoni (Wuhan isolate). A Changes in parasitemia of…

Human study The study was approved by the institutional review boards (Ethics Commission of the Medical Faculty of Technical University Munich (protocol 193/17S) and Ethics Committee of the Medical Faculty of Heidelberg University (protocol S-206/2011)) and all patients provided written informed consent under these protocols. The study was conducted in…

Bacterial Genome Sequencing in Bengaluru | ID: 23080056348 Product Specification Service Location Bangalore Minimum Order Quantity 1 Product Description Preparation of gDNA Library, 2×150 PEIllumina Hiseq sequencing to generate 1GBData. Additional Information Item Code 9992 Delivery Time 5-7 days Interested in this service?Get Latest Price from the seller Contact Seller…

Challenges in DNA Mutation Detection: Molecular diagnostics of cancer has underscored the significance of driver genetic alterations in tumor growth and metastasis1, 2. Although genetic variations hold abundant diagnostic and therapeutic clues for cancer patients, detecting mutations in cfDNA proves to be a major challenge due to their typically low…

Cyntia Gracesella Patintingan,1 Melva Louisa,2 Vetnizah Juniantito,3 Wawaimuli Arozal,2 Silmi Hanifah,1 Septelia Inawati Wanandi,4 Rajarajan Thandavarayan5 1Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia; 2Department of Pharmacology and Therapeutics, Faculty of Medicine, Universitas Indonesia, Jakarta, Indonesia; 3Department of Veterinary Clinic Reproduction and Pathology, Faculty of Veterinary Medicine, Agriculture Institute of Bogor,…

A study conducted at Taipei Veterans General Hospital showed metagenomic next-generation sequencing (mNGS) provided an efficient and accurate alternative to blood culture in identifying pathogens in patients with suspected sepsis. The researchers employed a workflow using a fractionation filter to remove host cells and enrich microorganisms, increasing sensitivity and lowering…

Primers, plasmids, bacterial strains, and growth conditions Primers, plasmids, and strains used in this study are listed in Tables S1–S3, respectively. Escherichia coli strains MG1655, DH10B, MDS42, and their derivates were cultured in LB medium at 37 °C, with shaking at 220 rpm. All the recombinants containing the temperature-sensitive pSC101-derived plasmid were cultured…

Clinical Study Finds the Company’s Devin™ Membrane and Metagenomic Assay More Accurate and Faster than Blood Cultures and Assays without a Host Depletion Filter Micronbrane Medical is dedicated to revolutionizing pathogen identification through easy, accurate and cost-effective metagenomic next-generation sequencing. — Dr. Mengchu Wu TAOYUAN CITY, TAIWAN, July 20, 2023…

A common ancestor of pathogenic Kingella species acquired the RTX toxin Consistent with existing literature, when the five Kingella species were assessed for β-hemolysis on BHI plates supplemented with 10% sheep blood, only K. kingae and K. negevensis were hemolytic (Fig. 1A), suggesting that the two pathogenic Kingella species produce the…

Patients The WGS and clinical data of 235 patients with A-T were provided by the Global A-T Family Data Platform of ATCP. Our access to the data was approved by the Data Access Committee of ATCP. Selected patients with A-T enrolled at the Manton Center for Orphan Disease Research under…

Isolate storage, growth, and identification Historical BFG isolates originally cultured from clinical material between 1973 and 2018 were stored either lyophilized or frozen in skim milk media at the National Institutes of Health Clinical Center Department of Laboratory Medicine (Bethesda, MD). Isolates were de-identified and metadata including year and source/site…

A global medical solutions company, that supplies bathing systems into the NHS, had a contamination outbreak at several UK sites. During routine water testing by 20/30 labs, Pseudomonas aeruginosa was isolated from the faucets and shower heads, but not the supply water. The baths were installed and tested prior to…

€0.00 Availability: Out of stock Availability: Out of stock Isolation of NGS quality gDNA (if required) with QC. Fragmentation with QC. Preparation of appropriate library for sequencing on Illumina platform with QC. Sequencing using relevant PE read chemistry to generate committed data. Sequencing Run information. Raw data information (number…

Identification and expression of GS genes in CHO cells Database analysis (see Supplementary information 1 indicated that the endogenous GS genes in CHO-K1 include two genes on chromosome 5 (GS5) and chromosome 1 (GS1), for which in-house RNAseq data and previous publication, listed in Supplementary Table S1, showed high and low…

Plasmid construction For U6+27 gRNA-expressing constructs, the DNA sequences including the hU6+27 promoter, a gRNA scaffold, and a gRNA insert site were synthesized and cloned into the pBluSKM vector. For Pre gRNA-expressing constructs, a second gRNA scaffold was inserted downstream of the gRNA insert site in the U6+27 gRNA-expressing constructs….

In an eight-year study of more than 600 community-dwelling older adults, Johns Hopkins Medicine researchers say they have further linked levels of cell-free DNA (DNA fragments resulting from cell death) circulating in the blood to chronic inflammation and frailty. The study is novel and expands on previous work, the investigators…

Newswise — In an eight-year study of more than 600 community-dwelling older adults, Johns Hopkins Medicine researchers say they have further linked levels of cell-free DNA (DNA fragments resulting from cell death) circulating in the blood to chronic inflammation and frailty. The study is novel and expands on previous work,…

Kingsmore SF, Cakici JA, Clark MM, Gaughran M, Feddock M, Batalov S, et al. A randomized, controlled trial of the analytic and diagnostic performance of singleton and trio, rapid genome and exome sequencing in ill infants. Am J Hum Genet. 2019;105(4):719–33. Article  CAS  PubMed  PubMed Central  Google Scholar  Costain G,…

Department of Dermatology, Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, People’s Republic of China Correspondence: Jianyou Wang, Department of Dermatology, Second Affiliated Hospital, Zhejiang University School of Medicine, 88 Jiefang Road, Hangzhou, 310009, People’s Republic of China, Email [email protected] Abstract: Lipoid proteinosis (LP) is a rare autosomal recessive…

1. Introduction Periodontitis is a chronic disease that affects 10% of the world’s population [1]. It is characterized by the interaction among bacterial, inflammation and genetic factors. Specific virulent oral microbials cause the host immune response in patients with a genetic predisposition. The inflammation of the periodontal tissues could result,…

Ethics The Australian Genomics Acute Care study has Human Research Ethics Committee approval (HREC/16/MH/251). Parents provided informed consent for participation in the study, following genetic counseling. Study design and participants The Acute Care Genomics program is a national multi-site study delivering ultra-rapid genomic testing to critically ill pediatric patients with…

Distinguishing SA from other inflammatory joint conditions is based on clinical and routine laboratory investigations representing a diagnostic challenge.4 The SF volume in normal joints varies from a few drops in small joints to several millilitres in larger joints. SF volume in the normal knee joint is from 0.2 to…

Statement on ethical considerations All animal work was approved and permitted by the Local Ethical Committee on Animal Experiments and conducted according to the Guidelines for Animal Experimentation recommendations (ARRIVE guidelines). In particular, mouse work related to C57BL/6NCrl mice was approved and permitted by the Institute of Molecular Genetics of…

I am trying to run MutSIgCV and got stuck with this error: MutSigCV allsamples.md.tc.ir.br.pr.ug.dbsnp.vep.maf \ “$anno”exome_full192.coverage.txt \ “$anno”gene.covariates.txt \ my_results \ “$anno”mutation_type_dictionary_file.txt \ “$anno”chr_files_hg19 ====================================== MutSigCV v1.4 (c) Mike Lawrence and Gaddy Getz Broad Institute of MIT and Harvard ====================================== MutSigCV: PREPROCESS ——————– Loading mutation_file… Error using MutSigCV>MutSig_preprocess (line 246)…

Introduction Phytophthora sojae is among the most destructive pathogens of soybean (Glycine max). An estimated annual worldwide loss of US$1–2 billion is caused by P. sojae, which was first identified as a soybean pathogen in the 1950s in Indiana, Ohio, and North Carolina (Schmitthenner, 1985; Tyler, 2007; Dorrance, 2018). The…

In a recent article published in the eLife Journal, researchers launched a possum excreta surveillance program across 350 km2 in the Mornington Peninsula near South Melbourne, Australia. The study aimed to gather data that could help predict future human Buruli ulcer (BU) case locations via statistical modeling. The 2022 outbreak…

Introduction Microalgae encompass a large number of organisms with both prokaryotic and eukaryotic nature, making them an archetypal platform for recombinant technology (Malla et al., 2021). The nutraceutical significance of microalgae is characterized by their rich natural biomolecules and high amount of protein, vitamins, and lipid substances (Potvin and Zhang, 2010;…

Introduction Perturbations in the infant gut microbiome during early life affect growth, development, and long-term health (Sherman, 2010; Carl et al., 2014; Tarr and Warner, 2016). Preterm infants have a physiologically and anatomically immature gastrointestinal tract that is known to be more permeable than that of term infants, and their…

Introduction Gitelman syndrome (GS) is an autosomal recessive disease, characterized by hypokalemic alkalosis, accompanied by hypomagnesaemia, hypocalciuria, low blood pressure, and hypocalcemia, first described by Gitelman in 1966.1 It is caused by mutations in the SLC12A3 gene, which is located on the long arm of chromosome 16(16q13) and encodes the…

Bacterial strains and cultivation Twenty-eight L. paracasei isolates (including L. paracasei Zhang) and a pglX gene-inactivated strain of L. paracasei Zhang were obtained from the Key Laboratory of Dairy Biotechnology and Engineering, Ministry of Education, at the Inner Mongolia Agricultural University of China. For strain activation, the bacteria were cultivated…

Plasmid design The information on each plasmid for combinatorial libraries is indicated in Supplementary Table S1. Plasmids used in this study to generate combinatorial libraries for SpCas9, enAsCas12a, CHyMErA, enAsCas12a (dual-gRNA), CHyMErA.v2, multiSPAS were deposited to Addgene (IDs: 189632, 189633, 189634, 189635, 189636, 189637, respectively). 3Cs oligonucleotide design The protocol…

scEC&T sequencing A detailed, step-by-step protocol of scEC&T-seq is available on the Nature Protocol Exchange46 and is described below. The duration of the protocol is approximately 8 days per 96-well plate. Cell culture Human tumor cell lines were obtained from ATCC (CHP-212) or were provided by J. J. Molenaar (TR14; Princess…

desgin a primer for CDS 0 I want to desgin a primer for the mRNA, now I have some cell samples, it contains gDNA, mRNA(not only the target gene) More detail: I need to use the pcr to amplyfy the target gene mRNA(mature), now I need to design a primer…

Recombinant protein expression and purification NanoLuc is the tradename of an engineered deep-sea shrimp luciferase variant developed by Promega Corporation36. Genes encoding fusion proteins were inserted into pET-21d(+) vectors between NcoI and XhoI sites. IDO-Gal3 genetic and protein sequences are provided in Supplementary Information. Plasmids confer ampicillin resistance and were…

Menasha J, Levy B, Hirschhorn K, Kardon NB. Incidence and spectrum of chromosome abnormalities in spontaneous abortions: new insights from a 12-year study. Genet Med. 2005;7(4):251–63. doi.org/10.1097/01.GIM.0000160075.96707.04. Article  PubMed  Google Scholar  Munné S, Chen S, Collis P, Garrisi J, Zheng X, Cekleniak N, et al. Maternal age, morphology, development and…

Introduction Rheumatoid arthritis (RA) is a chronic systemic autoimmune disease, mainly characterised by symmetrical pain and swelling of small joints in the hands and the feet.1 When chronic inflammation is not properly controlled over a long period of time, it can eventually lead to irreversible damage to the joint structures….

Plant material Streptocarpus grandis (RBGE lineage 19771210) and Streptocarpus kentaniensis (RBGE lineage 19951992) were cultivated in the glasshouses at the Royal Botanic Garden Edinburgh. DNA extraction method for Streptocarpus PacBio HiFi long-read sequencing To develop the protocol, small scale extractions were carried out with approx. 3 g leaf tissue as input…

Ethical approval, DNA samples and oligonucleotides All patients provided written informed consent to allow the collection of blood and/or tumor tissue and the analysis of clinical and genetic data for research purposes. The IRB of the Dana-Farber Cancer Institute and New York University Grossman School of Medicine approved these protocols….

. 2023 Apr 18;5(2):lqad035. doi: 10.1093/nargab/lqad035. eCollection 2023 Jun. Affiliations Expand Affiliations 1 Biology Department, Clark University, 950 Main Street, Worcester, MA 01610, USA. 2 Howard Hughes Medical Institute and Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720, USA. Free PMC article Item in Clipboard Robert A Drewell et…

Mosquitoes The Armigeres subalbatus GZ strain (Guangzhou Guangdong Province, China) was established in the laboratory in 2018 and reared in 30-cm3 nylon cages in the insectary at 28 ± 1 °C with 70–80% humidity and a 12:12 h (light: dark) light cycles. Larvae were fed with finely-ground fish food mixed 1:1 with yeast powder,…

Introduction Increasing antibiotic resistance is a worrisome trend being observed worldwide. Methicillin-resistant Staphylococcus aureus (MRSA) was first reported in the United Kingdom (UK) in 1961 and has since been reported throughout the world as a nosocomial pathogen.1 A steady increase in the occurrence of MRSA strains has been reported in…

Sequencing and assembly of mitochondrial DNA 1 Hi, I have just assembled the nuclear genome of two species and I am wondering if it is possible to also assemble the mitochondrial DNA using the same DNA reads I used for the nuclear genome. Is there a ways to “rescue” mitochondrial…

The latest issue of Nature Biomedical Engineering contains a study that reveals a new technique for safer gene editing. It reduces the problem of mutations in gene therapy. CRISPR/Cas9 is a technique that has been widely used for editing the genome. It allows you…

The TRACERx 421 cohort The TRACERx study (clinicaltrials.gov/ct2/show/NCT01888601) is a prospective observational cohort study that aims to transform our understanding of NSCLC, the design of which has been approved by an independent research ethics committee (13/LO/1546). Informed consent for entry into the TRACERx study was mandatory and obtained from every…

Ethics statement All animal handling and experimental procedures were approved by the Qufu Normal University Biomedical Ethics Committee (approval number: 2022001). Sample and sequencing Baer’s pochard tissue for whole-genome sequencing was obtained from a dead individual that had strayed into a fishing net in Shandong (China). The muscle tissue that…

Schematic of the experimental workflow employed in this study. A sample of gDNA extracted from each strain of each species was split in order to prepare parallel sequencing libraries for both long-read (PacBio RSII) and short-read (Illumina MiSeq) sequencing. Biological replicates for transcriptomics, proteomics, and metabolomics were generated from a…

Introduction Coronary heart disease (CHD) is the most common heart condition worldwide and the leading cause of death of elderly men and women (1). Despite recent declines in developed countries, both CHD morbidity and mortality continue to increase rapidly in developing countries. Various factors are involved in CHD pathogenesis, including…

Collection and maintenance of Drosophila biauraria We used laboratory stocks of Drosophila biauraria (Diptera; Drosophilidae), which were originally collected at the Field Science Center for Northern Biosphere, Hokkaido University located at Tomakomai, Hokkaido in 2011 and 2015 using standard banana traps and sweeping22. Females were brought into the lab and…

Steve Kanner, Ph.D. has been the Chief Scientific Officer of Caribou Biosciences since 2017, and is responsible for the company’s therapeutic discovery, research, and development activities. Steve received his undergraduate degree in Genetics from the University of California, Berkeley, and his Ph.D. in Immunology and Microbiology from the University of…

Below is listed the optimal sample requirements needed.  The stated volumes provide us with enough of the sample for our internal sample checks and library preparation.  Samples can be provided more concentrated than suggested and will be diluted as needed. Please contact your Sales representative for minimum sample requirements.  …

In this work, a rapid, sensitive, and portable detection method for Candidatus Liberibacter asiaticus based on recombinase polymerase amplification combined with CRISPR-Cas12a was developed. Huang long bing, abbreviated as HLB. It’s a devastating citrus disease worldwide. In the most citrus-producing countries, such as China and the USA, HLB is caused…

Ethics statement Informed consent for diagnostic and research-based studies was obtained for all subjects in accordance with the Declaration of Helsinki protocols and approved by local institutional review boards (Yorkshire & The Humber – Leeds Bradford Research Ethics Committee (13/YH/0310), the Sydney Children’s Hospitals Network Human Research Ethics Committee (HREC/10/CHW/113)…

Data-driven culturomics using phenotypes and automation Colony picking is a classic microbiology method for clonally isolating bacterial strains. Colony growth on plates depends on many factors, including the composition of the media (for example, available nutrients), atmospheric conditions (for example, level of oxygenation), presence of inhibitory molecules (for example, antibiotics),…

doi: 10.3389/fgene.2022.917993. eCollection 2022. Krinio Giannikou  1   2 , Katie R Martin  3 , Ahmad G Abdel-Azim  1 , Kaila J Pamir  4 , Thomas R Hougard  4 , Shefali Bagwe  4 , Yan Tang  4 , Jeffrey P MacKeigan  3 , David J Kwiatkowski  1 , Elizabeth P Henske  4 , Hilaire C Lam  4 Affiliations Expand Affiliations…