2 replies to this topic

[pcr_newbie]

Hello,
  I have a small amount of starting material (10ul) and need to conserve my use of it. I have an un-amplified PCR product from an unsuccessful PCR. Is it possible to re-use this unamplified product as starting material for a new PCR? If so, should I increase the amount of this starting material to use?

Thanks,

Cait

[ivanbio]

If your unsuccessful PCR did not amplify the product, then I would not use it. I would bite the bullet and use a small amount of your starting material and amplify it. Then save some of that and amplify from it again. That way you could use 0.5 ul of your starting material and amplify, and re-amplify, from it plenty of amplicon.

My two cents.

[Ameya P]

pcr_newbie, on 03 January 2011 - 08:16 AM, said:

Hello,
  I have a small amount of starting material (10ul) and need to conserve my use of it. I have an un-amplified PCR product from an unsuccessful PCR. Is it possible to re-use this unamplified product as starting material for a new PCR? If so, should I increase the amount of this starting material to use?

Thanks,

Cait

The template in the unsuccessful PCR is now highly diluted and even if your PCR were successful this time around, the amplified PCR product might not be enough to show up on the gel, and you would think the PCR is not working. This would result in wasting of resources and your time in trying achieve something which is not necessary. So, go ahead use the template, amplify your product and then there shall be abundance....