PickMutant™ DNA Shuffling Kit

PickMutant™ DNA-Shuffling Kit is based on the method Developed by Stemmer. Stemmer introduced DNA shuffling, the first homologous recombination method, in 1994.

DNA shuffling involves the digestion of a gene by DNase I into random fragments, and the reassembly of those fragments into a full-length gene by primerless PCR: the fragments prime on each other based on sequence homology, and recombination occurs when fragments from one copy of a gene anneal to fragments from another copy, causing a template switch, or crossover event. While randomly recombining the DNA sequences, the technique also introduces new point mutations at a relatively high rate (0.7%).
DNA shuffling consists of four steps: (i) preparation of genes to be shuffled, (ii) fragmentation with DNase I, (iii) reassembly by thermocycling in the presence of a DNA polymerase, and (iv) amplification of reassembled products by a conventional PCR.

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

Includes
- 40 μL Taq Polymerase (5 U/μL)
- 500 μL PCR Buffer (10x)
- 80 μL dNTP Mix (10 mM)
- 100 μL DNase I Solution (0.1 U/ μL)
- 200 μL Reaction Buffer (10x)
- 100 μL Stop Solution
- 3 x 1.5 mL PCR-grade Water

Applications
- Generation of a library of mutants.
- Slightly different genes coding for the same product.
- Homologous recombination of genes.
- Generation of gene libraries.
- Recombination of sequence blocks.
- In vitro Molecular evolution.

Shipping & Storage
Shipped in: Gel pack.
Storage: -20 ºC (NON Frost-Free Freezer) and Avoid freeze/thaw cycles.