PickMutant™ Random Mutagenesis Kit

PCR is routinely performed with standard Polymerases, yielding a final PCR product with few errors. Although Polymerases vary in fidelity, the inherent Mutation rate of PCR is insufficient to generate the high frequency of mutations necessary to generate a mutant library. The addition of nucleotide analogues to the PCR reaction reduces Polymerase fidelity increased substitution frequency in the PCR products.

PickMutant™ Random Mutagenesis (by Nucleotide Analog dPTP) Kit is based on the incorporation of Mutagenic dPTP, into an amplified DNA fragment by PCR. The Mutagenic dPTPs are eliminated by a second PCR step in the presence of the four natural dNTPs only, resulting in a rate of Mutagenesis of up to 20% (Zaccolo et. Al).

dPTP is an excellent substrate for Taq Polymerase (Km =22 mM versus Km = 9.5 mM for TTP); it is incorporated in place of TTP and, with a ≈fourfold lower efficiency, in place of dCTP. dPTP produces four transitions (A→G, T→C, G→A and C→T). Two transitions (A→G and T→C) occur at higher frequency than the other two (G→A and C→T) (Zaccolo et. Al).

Advantages & Features
- Highly Efficient.
- Reliable.
- Robust.

Includes
- 40 μL Taq Polymerase (5 U/μL)
- 200 μL PCR Buffer (10x)
- 100 μL dNTP Mix (10 mM)
- 100 μL MgCl2 (25 mM)
- 40 μL dPTP (10 mM)
- 2 x 1 mL PCR-grade Water

Applications
- Random Mutagenesis by dPTP Analog.
- Generation of a library of mutants.

Shipping & Storage
Shipped in: Gel pack.
Storage: -20 ºC (NON Frost-Free Freezer) and Avoid freeze/thaw cycles.
Advice: dPTP is most stable when stored as aqueous solution in the freezer (- 20° C necessary, – 80 °C recommended), however, at Ambient Temperature the compound slowly starts to decompose. Thus, in order to maintain its original high quality, it is recommended to allow thawing only before using the product. If you will not use up the vial with one application, please aliquot the contents of the vial in order to avoid repeated freeze/thaw cycles for the rest. When making such aliquots be sure to operate quickly and to freeze the vial again as soon as possible.