Manas Sanoria

6 Questions 2 Answers 0 Followers

Questions related from Manas Sanoria

Will the temperature of PCR lid affect the overnight ligation at 16 degree celsius?

I had set up overnight ligation reaction at 16 degree celsius in a PCR machine but the lid temperature was high around 95 degrees. Will it affect the efficeincy of ligation if done in PCR in this way?

23 January 2024 2,418 2 View

Is there any non kit based method to isolate PCR amplified product?

Is there any manual method to precipitate PCR amplified product?

02 December 2023 5,503 4 View

For doing blunt end cloning shall I dephosphorylate the linearized vector before blunting?

To ligate blunt insert into blunt vector does the vector need to be dephosphorylated at the time of ligation? I have digested my vector with restriction enzyme that create sticky ends that...

16 November 2023 412 0 View

Will the gene under EF1a promoter (from mammalian expression vector) be expressed in any bacterial host?

I have inserted a gene in downstream to Ef1a promoter of my mammalian expression vector. Can I check the protein expression using any bacterial expression host? Or, will it only express in any...

07 November 2023 3,372 4 View

What are the odds that, the amplicon generated using primers carrying overhangs will have blunt end and does not carry any overhangs ?

The DNA polymerase fills in the gap due to overhangs (using a pair of primers carrying overhangs) during the 2nd round of replication in PCR. Does those gaps actually get filled or they have...

03 November 2023 6,360 1 View

How can I ligate an insert digested with same restriction enzyme from both end to a vector digested with same enzyme?

I have insert having flanking regions with XbaI site and and a vector digested with XbaI. How can I prevent self ligation in insert as well as vector before ligation?

01 November 2023 8,833 1 View