Tag: pUC19

Methods Approval for animal experiments The prof. Dorota Ziyba-Przybylska as a Chairman of ethics review board and Dean of the Faculty of Animal Science, University Agriculture in Krakow considers that this type of project does not fall under the legislation for the protection of animals used for scientific purposes. The…

The main difference between pBR322 and pUC19 is that pBR322 has two origins of replication, one derived from the ColE1 plasmid and another from the pMB1 plasmid, whereas pUC19 has a single origin of replication derived from the pMB1 plasmid. pBR322 and pUC19 are both important plasmids in molecular biology….

Version Alpha Assignment #10 (50 pts) Name: Evelynn Najork *****type 2 Work that is done inside Lab 10 – DNA INames: Evelynn and Cady Without belaboring a repetitive comment, this assignment will SIGNIFICANTLY challenge you. We’ve gone from doing ‘ABC’ in Lab 8, to writing a book in Lab 9,…

Transcribed image text: Molecular Biology \& Genetics – MOB673 Independent Laboratory Project Functional Cloning of Tetracycline Resistance Markers in E. coli using Native Promoters – Plasmid Engineering \& Selection Aim: Functional migration of Tetracycline Resistance Marker from a donor plasmid to a recipient plasmid utilizing expression cloning and constitutive, native…

Falkowski P. G., Raven J. A. Aquatic Photosynthesis (Princeton Univ. Press, 2013). Sanchez-Baracaldo, P., Bianchini, G., Wilson, J. D. & Knoll, A. H. Cyanobacteria and biogeochemical cycles through earth history. Trends Microbiol. 30, 143–157 (2022). Article  CAS  PubMed  Google Scholar  Blankenship, R. E. Early evolution of photosynthesis. Plant Physiol. 154,…

Natalia Wood Genomic DNA Analysis through PCR and RFLP using Gel Electrophoresis April 11, 2022 Dr. Heidari Abstract: This course aimed to explore ancestral relatedness through the use of PCR and RFLP using gel electrophoresis, and a series of experiments were conducted to gain further insight on this topic. Initially,…

Transcribed image text: Pretend you are transforming mutated E colif with the pUC18/pUC19/plasmid that contains genes for a functional lac operon. Colonies were allowed to grow and MiniPrep was performed to isolate the DNA. You subject DNA to several different restriction enzymes and separate fragments with gel electrophoresis. Samples…

Overall structure of BPSL1038 The crystal structure of recombinant BPSL1038 (rBPSL1038) protein (~12 kDa) was determined by the single wavelength anomalous (SAD) dispersion method using the SeMet-BPSL1038 protein. The SeMet-BPSL1038 (smBPSL0138) and native BPSL1038 (rBPSL1038) crystals diffracted to 1.88 Å and 1.55 Å resolution, respectively. Both crystals belong to space group C2221 with…

Hi All, and thank you for taking the time to try and help. I am using this pipeline to calculate the coverage of my sequencing across: You will need: Example install, how all of the above are installed: ./configure –prefix=/Path/to/new/folder I just called put in ./configure –prefix=/Users/phr361/tool cd tools_folder make…

Use this tool to design a custom DNA oligonucleotide, with any of a variety of 3′ and 5′ modifications, at scales ranging from 25 nmole to 10 µmole. Select desired purification method (normal or reversed-phase chromatography, HPLC, or PAGE). Use this tool to design a custom DNA oligonucleotide, with any…

Bacterial strains, vectors and cloning E. coli XL-1 Blue (recA1 endA1 gryA96 thi-1 hsdR17 supE44 relA1 lac [F’ proAB lacI q Z\(\Delta \)M15 Tn10 (Tet r)]), originally purchased from Stratagene (USA), was used for all phage display selections and Fab expression in screening. pEB32x6 phagemid vector (later “display vector”) was…

pUC19-FrgA is the pUC19 (2686 bp) plasmid carrying fragment A with a length of 2 kb in the the SalI site. This fragment shall be isolated and cloned into the SalI site of yeast shuttle vector pRS (8.5 kb). You must show your calculations to obtain credit. Reagents: Plasmids are…

would have cloned the gene for red fluorescent protein RFP DsRed2 into the plasmidpUC19 and transformedE. coli DH5α.After selecting for clones, we would have isolated the proteins from these clones and the clonesisolated from the transformation experiment (Lab 8,E. coli that had the pGLO plasmid).A Western blot will be performed…

you decide you want to extract the pUC19 plasmid with the RFP gene to use in an upcomingexperiment.Your benchmate set up the overnight culture for you.They inoculated the clones in LB + IPTG.You took the cloudy culture and spun the cells down.You perform the plasmid extract and get a low…

Transcribed image text: 1. You discover a bacterium that produces a super lethal antibiotic. You also found the gene that encodes for the enzyme that makes this antibiotic. You call the gene for the enzyme sla. You want to purify large amounts of the antibiotic and bottle it for sale…

Transcribed image text: 8. What is the purpose of IPTG and X-gal in transformation experiments? Be detalled and specific for each reagent. Assume that the cells are transformed with pUC19. 9. You have performed a chemical transformation experiment in which a recombinant pUC19 plasmid has been added to competent cells….

In a recent study published in Nature Biotechnology, researchers developed a whole-genome sequencing (WGS) method that read four canonical deoxyribonucleic acid (DNA) bases, viz., adenine (A), cytosine (C), guanine (G), and thymine (T) plus 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC), epigenetic variants of unmodified C to yield an accurate six-letter digital…

Transcribed image text: QUESTION 1 During the procedure of cloning a gene of interest (GOI) in the pUC19 plasmid, both the vector and fragment to be inserted were treated with alkaline phosphatase following restriction enzyme digestion. After the ligation and transformation steps, no colonies were found on the agar plate….

In nature, microbes have evolved different systems to sense external stimuli. Synthetic biology approaches (1) repurpose these systems as biosensors to specifically and sensitively detect various targets of interest. Although various highly sensitive and specific laboratory-based analytical methods (including high-performance liquid chromatography and mass spectrometry) can detect small-molecule targets, they…

W. Hong et al. (12) previously demonstrated the utility of a CRISPR-Cas12a system in the low-GC Gram-positive organism C. difficile (12). As C. difficile and E. faecium have similar codon usage, we utilized Cas12a placed under the control of the tetracycline-inducible promoter (Ptet) from pRPF185 on the pMTL82151 backbone (pDL1,…

An efficient method for the transformation of Lactobacillus acidophilus TK8912 by electroporation is presented. The plasmid vector pULA105E was constructed by joining a cryptic plasmid, pLA 105, from L. acidophilus TK8912, the Escherichia coli vector pUC19, and the erythromycin resistance gene of the Streptococcus-Escherichia coli shuttle vector pVA838. Plasmid pULA105E…

Abstract Ribonucleoprotein (RNP) complex–mediated base editing is expected to be greatly beneficial because of its reduced off-target effects compared to plasmid- or viral vector–mediated gene editing, especially in therapeutic applications. However, production of recombinant cytosine base editors (CBEs) or adenine base editors (ABEs) with ample yield and high purity in…