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Contents

Purpose

Efficient cloning of PCR products.

Materials

Procedure

Reshma's procedure

(Note that the kit details a slightly different procedure which may give superior efficiencies.)

  1. Mix 1 μL PCR product (amplified with Platinum PCR supermix) with 0.5 or 1μL TOPO vector (0.5μL should be sufficient.)
  2. Incubate 5 mins on the bench top
  3. Place on ice
  4. Transform

Notes

Resources

References

Error fetching PMID 8780871:
Error fetching PMID 1658796:
Error fetching PMID 10756188:
Error fetching PMID 7798275:
Error fetching PMID 10207160:
  1. Error fetching PMID 8780871: [Brownstein-Biotechniques-1996]
  2. Error fetching PMID 1658796: [Shuman-PNAS-1991]
  3. Error fetching PMID 10756188: [Cheng-NAR-2000]
  4. Error fetching PMID 7798275: [Shuman-JBC-1994]
  5. Error fetching PMID 10207160: [Heyman-GenomeResearch-1999]
All Medline abstracts: PubMed HubMed

Patents

  1. US Patent 6,916,632; Methods and reagents for molecular cloning. (On topoisomerase-mediated cloning using a 5' overhang and blunt end.) pdf html
  2. US Patent 5,766,891; Method for molecular cloning and polynucleotide synthesis using vaccinia topoisomerase pdf html
  3. US Patent 6,653,106; Topoisomerase based ligation and cloning methods pdf
  4. US Patent 6,548,277; Method for molecular cloning and polynucleotide synthesis using vaccinia topoisomerase pdf html
  5. US Patent 5,487,993; Direct cloning of PCR amplified nucleic acids pdf html
  6. US Patent 5,827,657; Direct cloning of PCR amplified nucleic acids pdf html