6 replies to this topic

[kmad]

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

[almost a doctor]

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

[phage434]

Are you detecting or making this fragment?

[kmad]

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

Are you detecting or making this fragment?

I am making a fragment.

[almost a doctor]

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

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

Are you detecting or making this fragment?

I am making a fragment.

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.

[kmad]

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

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

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

Are you detecting or making this fragment?

I am making a fragment.

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.

Yes, It`s actually ssDNA a probe oligo.

[phage434]

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