Monika Jain

13 Questions 14 Answers 0 Followers

Questions related from Monika Jain

Sonication of protein sample?

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 1,433 9 View

How much concentration of Protease inhibitor cocktail should I use in buffer?

How to decide how much concentration of protease inhibitor cocktail is required in buffer for protein expression ?

11 May 2017 6,680 3 View

Query about purification of fusion protein by Ni NTA chromatography ?

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 9,301 7 View

Regarding linear and exponential PCR amplification during quick change and Inverse PCR site directed mutagenesis (SDM)?

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 3,266 2 View

Interpretation of gel before and after DpnI digestion?

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 10,030 6 View

Troubleshoot for quick change site directed mutagenesis. Will the nicked DNA be repaired in DH5alpha?

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 3,050 3 View

Regarding resuspension solution provided with Gene Jet miniprep plasmid Isolation kit ?

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 7,625 3 View

Regarding recombinant plasmid isolation through Kit?

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 3,677 1 View

What is control for DpnI digestion after quick change site directed mutagenesis?

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 7,719 8 View

What % of agarose gel should I use to separeate 500bp and 100 bp restriction enzyme digested DNA sample ?

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 3,221 10 View

Regarding viability of Ramos cell line. I had revived Ramos cell line one week ago but day by day cells are dying, how to troubleshoot this problem ?

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 188 3 View

If anyone knows then please tell me insert size capacity of PET vector ?

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 8,161 3 View

Which Antibiotic resistant Lb agar plate is required for rosetta competent cell preparation?

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 1,889 4 View