3 replies to this topic

[Schrodinger's Cat]

Hi guys,
I'm pretty new here but I like the look of this place, so sorry if I've broken protocol.

What I'm looking for is a good product that enables 96 well cleanup of PCR products for the purposes of sequencing.
I would also rather use a centrifugation step in this process rather than using a vacuum pump, any suggestions.
Cheers guys.

[Schrodinger's Cat]

Oh yeah my pcr products are from about 500bp to 900bp, if that information helps anyone to help me out.
Cheers.

[parasuis]

I use the NucleoFast 96 PCR by Macherey-Nagel (http://www.macherey-nagel.com)  and it works like a charm. I use their vacuum manifold, but you can also stick it in a centrifuge.

[wcousin]

I've used Zymo's 96 well pcr kit.  Here is a little info of the kit:

Quote

-High-Throughput PCR Clean-Up / Gel Extraction-

The 96-Well PCR Clean-Up/DNA Extraction Kits provide a high-throughput, rapid and economical method to purify DNA fragments. Chaotropic salt is used to denature enzymes and in this condition, DNA fragments are bound by the glass fiber matrix in each well of the plate. Once the contaminants have been removed, the purified DNA is eluted by a low salt elution buffer or water. Salts, enzymes, and unicorporated nucleotides are effectively removed from reaction mixtures without toxic phenol extraction or alcohol precipitation. This entire protocol can be completed in 30-40 minutes, and the eluted DNA is ready to use in restricted digestion, litigation, PCR and sequencing reactions.

KEY FEATURES:

• Sample Size: Up to 50ul of PCR product
  
• Format: 96-Well Plates
  
• Operations: Centrifuge / Vacuum Manifold
  
• Binding Capacity: 10mg per well
  
• DNA Size: 50bp - 10kb
  
• Recovery: 80-90% for Gel Extraction / 90-95% for PCR Clean-Up
  
• Operation Time: 30 minutes for PCR Clean-Up / 40 minutes for Gel Extraction