End-It™ DNA End-Repair Kit

End-It™ DNA End-Repair Kitは、5’、もしくは3'の突出末端DNAを5'-phosphorylationします。blunt-ended DNAは、プラスミドやコスミド、フォスミド、BACなどへのblunt-end cloningに使用できます。blunt-ended DNAへの転換は5'→3' polymeraseによる伸長とT4 DNA Polymeraseの3'→5' exonuclease活性により行われます。blunt-ended DNAのphosphorylation のためのT4 Polynucleotide KinaseとATPもキットに含まれます。End-It DNA End-Repair KitはEPICENTRE社pWEB™ Cosmid Cloning Kit、pWEB-TNC™ Cosmid Cloning Kit、EpiFOS™ Fosmid Library Production Kit、 CopyControl™ Fosmid Library Production Kitで提供されているEnd-Repair Enzyme Mixと同じものです。

End-It DNA End-Repair Kitは20反応分(repair of up to 100 μg of genomic DNA)の試薬が含まれています。end-repaired, 5'-phosphorylated DNAは、Fast-Link DNA Ligation Kitを使用することでblunt-ended, dephosphorylated cloning vectorへ効率的にライゲーションすることが可能です。

Application

Prepare sheared, nebulized, or restriction enzyme digested genomic DNA for blunt-end cloning into plasmid, cosmid, fosmid, or BAC vectors.

Benefits

The repaired DNA is blunt-ended and 5'-phosphorylated for immediate blunt-end ligation into plasmid, cosmid, fosmid, or BAC vectors.

High specific activity T4 DNA Polymerase and T4 Polynucleotide Kinase are included for complete conversion of 5'- or 3'-overhang DNA to 5'-phosphorylated, blunt-ended DNA.

Figure 1. DNA fragments containing any type of ends are rapidly and efficiently converted to 5'-phosphorylated, blunt-ended DNA using the End-It™ DNA End-Repair Kit.

The end-repaired DNA is ready for blunt-end ligation into the cloning vector of choice using, for example EPICENTRE's Fast-Link™ DNA Ligation Kit.

Kit Contents

• End-Repair Enzyme Mix including : T4 DNA Polymerase and T4 Polynucleotide Kinase
• End-Repair 10X Buffer
• dNTP Solution
• ATP

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