PCR Terminator® Kits

Optimizes cloning of PCR products using blunt vectors

Amplification of DNA by the polymerase chain reaction (PCR) requires the use of a thermal stable DNA polymerase. DNA polymerases can be categorized as either non-proofreading (i.e., without inherent 3'- exonuclease activity) or proofreading (i.e., with 3'-exonuclease activity). PCR products generated with non-proofreading polymerases such as Taq, Tth, or Tfl primarily have single-base adenosine orguanosine extensions at their 3´ termini. These products can be cloned using blunt vectors, like the CloneSMART®, pSMART® vectors with the PCRTerminator Kit.

Amplification products generated by proofreading polymerases such as Vent® or Pfu are primarily blunt-ended and can be cloned into blunt vectors directly without end repair. However, it is essential that the termini of the PCR fragment or the vector have 5' phosphate groups. Since vectors are commonly dephosphorylated to prevent self-ligation, the PCR product must be phosphorylated. The PCRTerminator Kit can also be used to repair these ends to allow cloning into dephosphorylated vectors, such as the pSMART® series.


 

PCRTerminator Advantages

Higher cloning efficiency. The PCRTerminator Enzyme Mix is optimized to produce the greatest percentage of blunt-ended, phosphorylated PCR products. As a result, ligation and cloning efficiencies are maximized, producing the greatest number of recombinants. Greater flexibility. The PCRTerminator Kit allows you to use any available blunt cloning vector, greatly increasing your options in selecting the best cloning strategy. Easy to use. Following a 30 minute incubation and a 15 minute heat-kill, simply purify the blunt PCR fragments and ligate into the vector of your choice.

PCRTerminator Applications



Cloning PCR products produced by any PCR enzyme

PCRTerminator Specifications



Each reaction end repairs 1 to 5 μg of amplification product for insertion into any blunt cloning site.
 

Click here for - PCRTerminator End Repair Kit Manual