• DNA/RNA Shuffling Kit
  • DNA/RNA Shuffling Kit

DNA/RNA Shuffling Kit

DNA shuffling, also known as in vitro homologous recombination of DNA molecules, is a directed evolution technique at the molecular level. DNA shuffling starts with one or more genes as starting materials, randomly breaking these genes into short DNA fragments, then performing PCR with mutual templates and primers (without external primers), and finally undergoing routine PCR (with external primers) to obtain PCR products containing a large number of DNA recombination mutations. DNA Shuffling K
$5.00
  • DNA/RNA Shuffling Kit

SPECIFICATION

                                                                          DNA Shuffling Kit

DNA shuffling, also known as in vitro homologous recombination of DNA molecules, is a directed evolution technique at the molecular level. DNA shuffling starts with one or more genes as starting materials, randomly breaking these genes into short DNA fragments, then performing PCR with mutual templates and primers (without external primers), and finally undergoing routine PCR (with external primers) to obtain PCR products containing a large number of DNA recombination mutations.
DNA Shuffling Kit
Ready to use, users only need to provide sample DNA templates without the need to prepare each component separately.
It can be used for DNA shuffling of homologous fragments with a length of about 1000bp.
The operation manual has been optimized to only generate DNA shuffling, while minimizing the probability of point mutations and saving a lot of optimization time.
This product is sufficient for 10 DNA shuffling reactions.
This product is only suitable for scientific research and cannot be used in clinical practice.
Component specifications
DNase I solution (1U/ μ L) 10 μ L
ten × DNA Shuffling special reaction solution A 100 μ L
ten × DNA Shuffling Special Reaction Solution B 100 μ L
two × DNA Shuffling specific primer free PCR MasterMix 1mL
two × DNA Shuffling specific primer PCR MasterMix 1mL
Ultra pure water 1mL
Storage: -20 ℃, valid for 1 year.
Mutated gene fragments, DNA Shuffling primers (second round PCR primers), and gel recovery kits.
Start 37 ℃ and 90 ℃ water or metal baths in advance.
1、 Preparation of mutated gene fragments
The mutated gene fragment can be DNA from multiple species containing homologous gene fragments. These gene fragments can be prepared by PCR or restriction endonuclease digestion.
The total length of these DNA fragments containing homologous gene fragments (or a portion of homologous gene fragments) should not exceed 1kb, and they are 200-400bp longer than the final product of DNA shuffling (the second round PCR product), as the primer position of the second round PCR is 100-200bp on the inner side of the template DNA prepared in this step.
Each DNA shuffling experiment requires 2-5 μ G Starting DNA fragment. If several homologous gene fragments are used, the ratio is 1:1:1, and the total amount is guaranteed to be 2-5 μ G.
The starting template DNA must be purified through gel recovery in order to completely remove residual plasmid templates, primers, and non-specific amplification products. If the primer in this step is not removed, it will cause serious interference to subsequent PCR.
Determine the template DNA concentration by measuring OD260. This solution is a homologous gene DNA template and is placed on ice for use.
2、 Enzymatic digestion and recovery of template DNA
Preparation 10 μ L DN
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