Looking for anyone with experience in assessment of differences between linear construct design versus plasmid based, STARR-Seq method...
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4690393/?
Please see Comparative analysis of enzymatically produced novel linear DNA constructs with plasmids for use as DNA vaccines
Adam A. Walters, PhD, Ekaterina Kinnear, MSc, [...], and John S. Tregoning, PhD
Genomic DNA is digested with a restriction enzyme, end labeled with biotin-dNTP with TdT, and then immunoprecipitated with Protein G Dynabeads coated with anti-biotin antibody.
11 December 2014 10,017 15 View
The subsequent step with Klenow exo- is done at 37 degrees to adenylate the blunt ended fragments. End repair uses a cocktail of T4 DNA polymerase, Klenow, and polynucleotide kinase.
11 December 2013 7,243 7 View
I have reverse sequences (AB1 format), can I base on reverse DNA sequences to perform nucleotide alignment, convert nucleotides to amino acids and deposit the sequence in GenBank database?
11 August 2024 5,138 1 View
I'm cloning a fragment of 3200 nts into plasmid. The cloning was successful, however, 02 amino acids were mutated. Now I want to fix these 02 aa by site-directed mutagenesis technique using...
08 August 2024 4,645 2 View
I am currently working on LncRNA; to know the lncRNA-protein interactions I want to do RNA pull down assay, so I need to design primers with T7 promoter. I need assistance in this regard.
07 August 2024 6,622 1 View
Hi all! My thesis groupmates and I are working on purifying an E. coli bacteriophage. The phage was handed down to us by the previous thesis group, but they only managed to purify it to about...
06 August 2024 5,801 4 View
Hi all, I need to introduce an ARS (autonomously replicating sequence) in my plasmid but I'm not sure which position would be the best. Does anyone have any suggestion? A picture of the plasmid...
05 August 2024 1,573 4 View
I want to introduce a point mutation (change in one nucleotide) into my gene of interest (DNA binding domain) I have designed primers as recommended on the Data sheet of the kit : -Both primers...
05 August 2024 9,059 3 View
I aim to be as skeptical as possible regarding whether a pair of orthologous genes results in the same phenotype in their different but related bacterial organisms under similar environmental...
05 August 2024 6,787 4 View
I am performing ligation of the plasmid and a target gene. The steps I have taken are: 1. Double digestion of the plasmid and target gene 2. Ligation of the plasmid with the target gene 3....
05 August 2024 2,570 3 View
TEP presentation caption (The Environmental Project) Re: Why should Washington’s DC, or any country government point of location think of as nowadays of as to being 'tomorrow as to come! if it...
03 August 2024 2,484 1 View
I have been attempting to extract DNA from Bacterial, Fungal and Yeast banked samples (>1e7 cells) using Prepman Ultra reagent and I seem to be struggling to obtain a sequence. Although the...
01 August 2024 2,079 0 View