Sanjay Premi

8 Questions 32 Answers 0 Followers

Questions related from Sanjay Premi

How can I extract biotinylated DNA from Agarose Gel?

Hi, I am trying to remove unligated, biotinylated linkers/adapters from my DNA by running a low melting point agarose gel (0.8%). After cutting the DNA out, and leaving the unligated small sized...

07 July 2015 4,124 4 View

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...

10 October 2014 5,164 5 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...

09 September 2014 1,497 5 View

How can I best carry out sequencing small PCR amplicon (~70 bp) by Sanger sequencing?

I am trying to sequence a 70bp PCR amplicon by Sanger Seq., using the PCR primers as sequencing primers. Will the small size of template be a problem? I tried it once, no result. Should I clone it...

08 August 2014 9,072 7 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...

06 June 2014 5,119 6 View

Does anybody know of a technique to detect the protein carbonyl adducts?

Oxyblot from Millipore can detect carbonylated proteins/amino acids, but what I am looking to do is detect a carbonyl compound added to a protein so that the actual carbonyl group is no longer...

04 April 2014 6,506 1 View

Does anybody work with the restriction enzyme Mme1?

I have tried all sorts of concentration variables of SAM, Mme1, and the target DNA. It works well on the pUC19 plasmid which has two Mme1 sites but gives 0% activity on a 100 bp PCR fragment...

03 March 2014 1,957 5 View

Has anybody used IgM as primary antibodies for immunofluorescence?

I am trying to see if the antigen is on the nuclear surface or inside the nucleus, but the antibody is giving non-specific background, even in the cells which are not supposed to have this...

11 November 2013 3,518 3 View