PCR amplifying 50bp ssDNA ? - PCR amplification (Nov/10/2009 )

Hi,

I need to amplify small 50bp ssDNA using PCR. How to design primers fitting in this small 50bp region ? Any ideas .. Highly appreciate it.

Thanks,

kmad

Could you just desing a forward and reverse 50bp primers and anneal them ?

Are you detecting or making this fragment?

phage434 on Nov 11 2009, 07:16 AM said:

I am making a fragment.

kmad on Nov 11 2009, 03:05 PM said:

I've just realised one thing:

on your first post you said 50bp ssDNA, did you mean dsDNA (as I assumed and misread), or did you actually mean single stranded DNA.

If the latter, isnt a 50bp ssDNA just an oligo?

If dsDNA, I still think you can hybridise 2x 50bp complementary oligos.

almost a doctor on Nov 11 2009, 09:00 AM said:

Yes, It`s actually ssDNA a probe oligo.

You order 50 bp oligos from IDT to do this. The major challenge is adding water to the tube (actually, I'd recommend TE).