2 replies to this topic

[uioli]

I run my PCR with primers from a well-known company.
But when I cloned PCR product, then Sequense it for 4 times from both ends of it. I found the primer have one extra base, which will change my DNA's frame.
Color electropherograms clearly showed an extra high peak there.
I have contact with the company, they said they will check the primer by "mass spec". Is there any other reliable way to check it? Whick company could do it?

[sbidw]

Dodgy primers are nothing new, normally you get N - 1 products though not extra bases. Even the big companies like MWG and Gibco, send out rubbish from time to time and there is nothing really that you can do about it. If you want to be more condident of getting a decent oligo most places will allow them to be PAGE purified, though this costs a lot more.  
If you cant be bothered to reclone the gene, just take the base out using quik change mutagenesis. At the end of the day compared to everything else in the lab oligos, are cheap, I would just order it again, and encourage the company to sort out their quality control.

[RichardColes]

I had a problem where the tube was labelled differently from the sheet. In this case I called up Gibco and they checked the sequence with me over the phone and then express delivered the correct oligo the next day. Just get them to do this, it cost nothing cos it was their fault.