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Questions related from Monika Jain
For the expression of subtle Nus tag fusion protein I have optimized sonication for 5ml culture, 50 percent amplitude, 1pulse off 1pulse ON for 30 second. With repeated cycles with the gap of...
21 June 2017 1,471 9 View
How to decide how much concentration of protease inhibitor cocktail is required in buffer for protein expression ?
11 May 2017 6,723 3 View
I am facing problem during purification of cytosolic fusion protein, by using Qiagen kit spin column (binding capacity 300 ug of protein) 80% protein is coming out in flow through so less yield is...
11 May 2017 9,368 7 View
During Quick change SDM F and R primers are complementary of each other when performing independent Forward and reverse primer PCR reaction (In different tubes) for quick change SDM then it is...
20 November 2016 3,318 2 View
I have performed Quick-change site directed mutagenesis by using stratagene protocol (Not kit). Mutagenesis primers were exact complementary and I have performed 1.Independent F & R...
17 October 2016 10,079 6 View
I want to create mutated DNA by using quick change site direct mutagenesis. Quick change works by using a pair of complementary primers with a mutation. In round of PCR cycles these primers anneal...
06 October 2016 3,104 3 View
I need to design primer for inverse PCR site directed mutagenesis where primers are not complementary to each other and anneal back to back at 5' end (not a single base is complementary in...
20 September 2016 6,667 1 View
Our lab is using the Gene jet miniprep kit for plasmid Isolation. Somehow, the resuspension solution or alkaline lysis I buffer or Buffer p1 is going to empty soon even though we still have a lot...
18 September 2016 7,690 3 View
I have isolated recombinant plasmid using all buffer solutions from GeneJet Mini prep plasmid isolation kit except resuspension solution. I used resuspension solution of promega Midi prep kit. I...
08 September 2016 3,729 1 View
I have cloned my insert DNA into pBluescript SK+ vector, after this cloning I have performed site directed mutagenesis by using PCR then digested it by DpnI digestion. what control should I use...
01 September 2016 7,784 8 View
I have 600bp long DNA, digested with Restriction enzyme to create 2 fragments one is 500bp long and another is 100bp long I want to see both the bands, so what percentage of agarose gel should I use?
12 July 2016 3,275 10 View
one week ago I had revived Ramos cell line in 10 cm plate by using RPMI 1640 Ramos medium. According to protocol, Ramos media should be 4500mg/L but I used 2000mg/L glucose. Next day I changed...
11 June 2016 248 3 View
I want to clone AID enzyme in expression vector so please suggest me which vector will be suitable for this purpose ?. I am planning to clone AID ( Size approx 4000 bp ) by using PET expression...
29 April 2016 8,190 3 View
I have been going through this paper: http://www.ncbi.nlm.nih.gov/pubmed/14724175 And it is unclear to me what exactly is meant by a "sterile transcript" and how it has been described in the...
09 July 2015 389 1 View
I want to confirm is there any antibiotic resistant Lb agar plate is requires for streaking of rosetta cells for rosetta comp. cell preparation OR I can use only Lb agar plate for this purpose. I...
01 January 1970 1,936 4 View
