Bhakti Jayant Vyas

6 Questions 5 Answers 0 Followers

Questions related from Bhakti Jayant Vyas

Does anyone use DSMZ institute's PCR method for deteting mycoplasma contamination in cell culture?

This method seems to be easy, sensitive and cost effective. All I am looking for internal control DNA. Also, please suggest any other easily available PCR methods or commercial kits

02 February 2019 9,076 2 View

How to detect biallelic mutation using T7 endonuclease assay?

I have transfected mouse ES cells with single gRNA, Cas9 system. I am able to detect cleaved bands after T7 endonuclease. However, how to determine if this is monoallelic or biallelic mutation?

12 December 2017 896 4 View

What should be the pulse length of electroporation for transfecting mammalian cells?

I am currently optimizing electroporation conditions for mouse embryonic stem cells. What should be ideal range of pulse duration?

07 July 2017 8,604 1 View

Has anyone used Biorad electroporation system to electroporate mouse ES cells E14Tg2a?

I wish to know electroporation conditions especially for transfecting Cas9 RNA and protein

12 December 2016 2,504 2 View

Is it a good idea to do an overlap extension PCR with KAPA polymerase?

I have tried amplifying my individual fragments using Phusion but it is not working consistently but always works fantastically with KAPA. I think I am unnecessarily wasting time on Phusion....

06 June 2016 9,321 4 View

What should be the ideal ratio of primer to template DNA in PCR?

I have tried doing a PCR with kapa as well as Phusion HF polymerase. I am getting amplification with kapa, but Phusion is just not working. Both the master mixes use same: 1.5mM MgCl2 and 0.2mM...

05 May 2016 6,935 3 View