Yue Ban

3 Questions 9 Answers 0 Followers

Questions related from Yue Ban

How can I decide the amount of cDNA added in PCR reaction if your gene of interest is in low abundance?

I am having trouble in PCR to amplify a low abundant gene. cDNA is made from total RNA. I was always adding 2 ng/ul - 4 ng/ul cDNA in the PCR reaction. Positive control like GAPDH always shows up...

31 August 2015 3,574 6 View

How can I do overlap PCR?

I need to clone a 4kb gene. I have problem in cloning the whole gene in a single PCR reaction. So I decided to amplify two fragments then overlap the two fragments in a another round of PCR. Both...

17 April 2015 4,496 3 View

Why has PCR amplifying 4kb DNA using cDNA as a template failed?

I used the SMARTer RACE kit from clontech to get the 3' and 5 ends of my target gene. Then I tried to PCR amplify the whole gene. I tried both the platinum Taq High fidelity DNA polymerase and...

26 January 2015 5,853 13 View