6 replies to this topic

[roshanbernard]

hi,
i have a gene which is 32 bp...can i use the pcr and produce the multiple copies it or is it too small for the PCR to do it...wat is the minimum gene length for the PCR....

[HomeBrew]

Why not buy two complementary primers that cover the gene in its entirety, and also leave appropriate overhangs so you can clone it? Mix the primers, melt, and cool slowly, then ligate this now-double-stranded DNA into an appropriately digested vector.

[phage434]

When you do this, don't forget to either order the oligos with 5' phosporylation, or kinase the annealed product to add 5' phosphates. They won't ligate with one or the other.

[T C]

Hey,

The lowest I have gone is 100 bp but why don't you just order a commercial olgo (in yr case gene) of 32 bp.

Best,
TC

roshanbernard, on Apr 13 2009, 07:22 AM, said:

hi,
i have a gene which is 32 bp...can i use the pcr and produce the multiple copies it or is it too small for the PCR to do it...wat is the minimum gene length for the PCR....

[HomeBrew]

phage434 is right -- I forgot to mention that you must have the oligos synthesized with 5' phosphorylation.

[hanming86]

32bp .. just do a gene synthesis. they have this service nowadays. they synthesize it and send it in a plasmid for ya.

[roshanbernard]

thanks a lot everybody...i think i will order for the gene with 5' phosphorylation,....