Bidur Paudel

3 Questions 8 Answers 0 Followers

Questions related from Bidur Paudel

How can I produce recombinant protein from E. coli without introducing mutations in my gene of interest?

Hello friends, I ligated my gene of interest in pET vector and transformed into BL21 cells and confirmed my insert through colony PCR. Further, I picked those PCR confirmed colonies for...

25 January 2015 4,022 12 View

Is anyone familiar with cloning Full length cDNA?

I try to clone entire Open Reading Frame of one of the proteins that I am studying and designed two sets of primers from 5'UTR and 3'UTR ( one I designed and one I got it from NCBI database)....

07 October 2014 9,778 4 View

How do I confirm if my site directed mutagenesis worked after transformation directly from colonies before sending it to sequencing?

I have 7.0 kb plasmid (with the insert). I performed Site directed mutagenesis and I have nice colonies grown in a plate (km). Now how do I confirm from the colonies that they contain the desired...

26 July 2014 8,805 8 View