BioForum > PCR amplifying 50bp ssDNA ?

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BioForum > Methods Discussion > PCR, RT-PCR and Real-Time PCR

kmad

Nov 10 2009, 01:19 PM

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

Nov 11 2009, 01:38 AM

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

phage434

Nov 11 2009, 06:16 AM

Are you detecting or making this fragment?

kmad

Nov 11 2009, 07:05 AM

QUOTE (phage434 @ Nov 11 2009, 07:16 AM)

Are you detecting or making this fragment?

I am making a fragment.

almost a doctor

Nov 11 2009, 08:00 AM

QUOTE (kmad @ Nov 11 2009, 03:05 PM)

QUOTE (phage434 @ Nov 11 2009, 07:16 AM)

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

Nov 11 2009, 09:09 AM

QUOTE (almost a doctor @ Nov 11 2009, 09:00 AM)

QUOTE (kmad @ Nov 11 2009, 03:05 PM)

QUOTE (phage434 @ Nov 11 2009, 07:16 AM)

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

Nov 11 2009, 02:23 PM

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

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