仕組み Pyrococcus species GB-Dポリメラーゼは、3'-5'エキソヌクレアーゼ活性を持つプルーフリーディング酵素です。この酵素をTaq DNAポリメラーゼと混合することで、忠実度が高まり、単純なDNAテンプレートから複雑なDNAテンプレートまで、広い範囲のターゲットサイズを増幅することができます。
• AccucPrime Taq DNAポリメラーゼ、高忠実性(5 U/μL)、1×200 μL • 10X AccuPrime PCRバッファーI、4×1.25 mL • 10X AccuPrime PCRバッファーII、4×1.25 mL • 50 mM MgSO4、1×1 mL
-10℃~-30℃で保存。
よくあるご質問(FAQ)
My oligonucleotide does not appear to be the right length when I checked by gel electrophoresis. Why is this?
Oligos should be run on a polyacrylamide gel containing 7 M urea and loaded with a 50% formamide solution to avoid compressions and secondary structures. Oligos of the same length and different compositions can electrophorese differently. dC's migrate fastest, followed by dA's, dT's, and then dG's. Oligos containing N's tend to run as a blurry band and generally have a problem with secondary structure.
The primers I am using worked for PCR initially, but over time, have stopped working. What happened?
Primers should be aliquoted for single use before PCR set-up. Heat just the aliquoted primers to 94 degrees for 1 min. Quick chill the primer on ice before adding to the PCR reaction. Some primers may anneal to themselves or curl up on themselves.
I don't see a pellet in my oligo tube order. Should I ask for a replacement?
The drying method dries the primer in a thin layer along the sidewalls of the tube instead of the bottom, therefore a pellet is not always visible and should still be ready to use.
There is a ball-shaped pellet at the bottom of my oligo tube. What is this and can I still use my oligo?
If the oligo was overheated, it will appear as a ball-shaped pellet attached to the bottom of the tube. This should not affect the quality of the oligo, and the oligo should be readily soluble in water.
There is a green color in my lyophilized oligo. Can I still use it?
If an oligo appears green in color, this is most likely due to ink falling into the tube. The oligo should still be fully functional. The color can be removed by doing an ethanol precipitation.
A rapid and general assay for monitoring endogenous gene modification.
Authors:Guschin DY, Waite AJ, Katibah GE, Miller JC, Holmes MC, Rebar EJ
Journal:Methods Mol Biol
PubMed ID:20680839
The development of zinc finger nucleases for targeted gene modification can benefit from rapid functional assays that directly quantify activity at the endogenous target. Here we describe a simple procedure for quantifying mutations that result from DNA double-strand break repair via non-homologous end joining. The assay is based on the ... More
Generation of adult human induced pluripotent stem cells using nonviral minicircle DNA vectors.
Human induced pluripotent stem cells (hiPSCs) derived from patient samples have tremendous potential for innovative approaches to disease pathology investigation and regenerative medicine therapies. However, most hiPSC derivation techniques use integrating viruses, which may leave residual transgene sequences as part of the host genome, thereby unpredictably altering cell phenotype in ... More