Farnaz Yusuf

6 Questions 13 Answers 0 Followers

Questions related from Farnaz Yusuf

Low intensity of my amplicon?

I am using primers having 60bps of overhangs in addition to the actual annealing region of around 22nts, thereby making my primers some 80-82nts long. When I am using these primers I get right...

06 June 2017 6,547 2 View

Can I use gel extracted PCR product as template for PCR using same primers and PCR conditions??

My expected amplicon size is 7.6 Kb. I am getting a very faint band of this size along with other non specific bands. I have tried every possible condition to increase the intensity of my band but...

05 May 2017 3,533 11 View

I have a plasmid with PBR322 origin, ARS CEN and f1 origin. I am curious to know whether f1 origin will have any effect?

I have a plasmid with PBR322 origin, ARS CEN and f1 origin. I am curious to know whether f1 origin will have any effect? I want to be sure it does not effect my expression.

06 June 2016 5,787 3 View

I am trying to clone a 4.2Kb insert in 5.7Kb vector both double digested with KpnI and sacI. But i am not able to get any positive clone.

The clonies i get are usually empty without any insert. I have tried 1:1,1:3,1;4 and 1:5 molar ratios so far.

02 February 2016 8,416 4 View

What possible reasons are there for having non-functional His-tagged nitrilase protein?

The protein expression is good. I get the expected protein size but it has no nitrilase activity. What's the problem?

11 November 2013 1,026 4 View

His-tag recombinant protein does not work in my functional assay - can anyone help?

I have recently expressed recombinant nitrilase in BL21. After purification I got a single band on SDS-PAGE of desired size. I stored the protein in 10% glycerol at -80C (elution buffer had 50mM...

11 November 2013 4,902 15 View