Stratagene Quikchange

Mutagenesis Kit

By: - Science - September 21, 2011
stratagene quikchange mutagenesis kit

The Stratagene QuikChange, or the QuikChange Mutagenesis kit has proven to help regarding DNA, being the most rapid, neat and efficient means of doing this.  Using the pfu Turbo in polymerase form, the kit replicates a template of the DNA, which has been taken from mutagenic nucleotide filled primers.  Seemingly straight forward, the kit creates an amplification that is equal to 2n as daughter DNA is unable to be used for PCR rounds templates.

Strategnic Quikchange uses nanograms of these templates because only 16 PCR cycles are needed to generate product mutations.  This means that it is easy to inspect the mutations on any agarose type of gel.  A successful type of mutagenic occurrence can be observed when additional bands are compared to template and other controlled types of reactions.

Primer sizes increase when the primer and amounts of nucleotides are mismatched.  In other words, a larger primer has to be utilized for larger amounts of mismatches.  Template to primer binding is best increased by maintaining high primer levels, helping in the compensation of primer dimers, which is totally unavoidable.  Most importantly the QuikChange Mutagenesis kit elegantly, efficiently and simply removes parental type templates by the resulting digesting Dpnl reaction.  This frequently used cutter that restricts enzymes can consume only DNA that is template and methylated.  As such the Dpnl buffer efficiently works, requiring no further purification at this point of the DNA.  As such the kit has sown to have good success types rates, working 1,2 and 3 nucleotide changes and providing a good extension of twenty-five base DNA pairs.  Additionally, the kit helps generate vast libraries of 2 codons, being separated by a single codon that has not yet been mutated, being randomly set into 50 base DNA pair primers.  However, issues surrounding the mismatching of base pairs that were no doubt somewhere in the DNA that was being used to transform certain bacteria, this was not seen as an issue, mutations are simply mutations and the repair systems of bacteria can easily correct any mismatching.

The Stratagene QuikChange, specifically the QuikChange Mutagenesis kit cannot be used for complete success, using similar protocols and individualized components found in the kits.  Despite this, the kit is seen as very helpful and if there are any issues, the customer service staff are very helpful, understanding exactly how this product works in relation to DNA work.

Photo: DNA – public domain photo – http://en.wikipedia.org/wiki/File:ADN_animation.gif


* * * * * 1 vote


Related Videos

 


Comments are closed.

  •