描述
TOPO® TA Cloning® Kits are designed for cloning PCR products directly from a PCR reaction in just 5 minutes (1). They use a pCR™-TOPO® Vector with covalently bound topoisomerase I for fast cloning and recombinants. pCR™-TOPO® Vectors include:
• 3´-T overhangs for direct ligation of
Taq-amplified PCR products
• Choice of T7 (pCR™2.1-TOPO®) or T7 and SP6 (pCR™II-TOPO®) promoters for
in vitro RNA transcription and sequencing. Each vector also contains M13 forward and reverse primer sites for sequencing.
•
EcoR I sites flanking the PCR product insertion site for easy excision of inserts
• Kanamycin and ampicillin resistance genes for your choice of selection in
E. coli
• Easy blue/white colony screening for selection of recombinants
TOPO® TA Cloning® Kits are also available in
combo kits combined with a PureLink™ Quick Plasmid Miniprep Kit (50 preps) for fast plasmid purification of your TOPO®-cloned inserts for downstream analysis.
内容及储存
Box 1:
• Topoisomerase I-activated pCR™2.1-TOPO® vector
• PCR buffer
• Salt solution
• dNTPs
• Control template
• M13 forward and reverse primers
• Control PCR primers
• Sterile water
Store at -5 to -30°C.
All reagents are stable for 6 months when properly stored.
Box 2:
• One Shot® Chemically Competent or Electrocomp™ E. coli
• S.O.C. medium
• Supercoiled pUC19 control plasmid
Store at -68 to -85°C.