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Questions related from Vishal Srivastava
I am trying to purify one of my protein of interest having molecular weight around 52 Kda. Upon over expression it comes in inclusion body. So I isolated and washed inclusion body and then...
16 August 2017 8,762 8 View
Dear all, I am planning to clone a gene in pET24a(+) Series vector but before going on for cloning i tried to sequence pEt24a(+) null vector from our lab repositiory, but in...
01 February 2017 6,973 3 View
I am trying to purify protein having molecular weight ~50Kda under denaturing condition through Ni-NTA chromatography and got following profile. Any suggestion to stop protein coming in flow...
20 June 2015 7,583 15 View
My cloned gene in pET23b contains a C-terminal his tag and mature form of protease. Its not getting expressed in e.coli cells. Even after transformation very little colony forms ~1-5 colonies...
21 February 2015 2,799 7 View
I successfully cloned a gene and checked it by colony pcr and restriction digestion. There after I transformed the construct in E.coli DH5alpha for plasmid prep and isolated sufficient amount of...
18 September 2014 10,038 9 View
Hello everyone, I prepared my vector (pET23b) and got 10.5ug of Vector dissolved in 150ul of nuclease free water and the insert is about 11 ug in 100ul. i wanted to perform a double digestion on...
29 July 2014 381 4 View
I had done PCR amplification of my gene of interest using bacterial genomic dna as template. Further gel extraction (thermo scientific) of 200ul pcr reaction volume had given pcr product having...
26 June 2014 4,375 16 View
