Sanjay Premi

4 Questions 20 Answers 0 Followers

Questions related from Sanjay Premi

Can DMSO in the PCR make amplicon size shift on agarose gel?

I expect a 70 bp amplicon which shows up only with 2% (or higher) DMSO in the PCR. But with DMSO, the size of this amplicon goes from 70 bp to ~110b bp on 3% agarose gel. Same happens when a...

03 October 2014 446 4 View

Is there a good protocol for making blunt ends out of denatured DNA using random hexamers and Klenow (Large Fragment) from NEB?

I am trying to make blunt ends out of nicked plasmid DNA using random hexamers and Klenow polymerase (large fragment) from NEB which has 3'-5' exo but no 5'-3' exonuclease activity. My goal later...

03 September 2014 3,846 4 View

T4 Endonuclease V is not cutting the UV exposed plasmid - any thoughts?

I am trying to cut ~ 5ug of pUC19 plasmid, exposed to very high UVC dose (~800 J/m2) using T4 endonuclease V from NEB. Everything is being done according to NEB Protocol. I tried increasing the...

11 June 2014 3,961 5 View

What might be causing a very high 260/280 ratio after Phenol:Chloroform extraction of DNA?

I am getting very high 260/280 ratios after a phenol chloroform extraction of small sized DNA fragments. Extra care was taken not to leave any phenol or chloroform in the DNA solution before...

12 May 2014 423 6 View