RECOMBINEERING

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• Background info
  • Summary
  • What is recombineering?
  • Requirements for recombineering.
  • Discovery of recombineering
  • Recombineering with dsDNA.
  • in vivo cloning
  • Recombineering with ssDNA
  • Site Map
  • Optimizing recombineering with ssDNA
  • On the mechanism of ssDNA recombination
  • Moving recombineering to other systems
  • “Recombinase”-independent recombineering
• Frequently asked questions
  • FAQ 1: How much antibiotic should I use for chromosomal/BAC (single-copy) or plasmid (multi-copy) constructs?
  • FAQ 2: What sequences do I use to make a drugR cassette for knock-outs?
  • FAQ 3: What happens if I grow the recombineering cells at temperatures higher than 32°C?
  • FAQ 4: Can’t I use an air shaker for the 42° induction?
  • FAQ 5: Do the oligos I order need to be purified?
  • FAQ 6: What happens if I induce for longer or shorter times?
  • FAQ 7: Why don’t I get any recombinants?
  • FAQ 8: I get tons of recombinants but when I analyze them, none are in the right place. What is wrong?
  • FAQ 9: What conditions should I use for electroporation?
  • FAQ 10: Why do my electroporations “pop” (arc)?
  • FAQ 11: Is the amount of NaCl in the LB medium important?
  • FAQ 12: How should I maintain the recombineering strains?
  • FAQ 13: What do I do with the tube containing the “gel-like stuff” in it when you send me a strain or plasmid?
  • FAQ 14: Why don’t I get the high frequencies that you report for ssDNA oligo recombineering?
• Strains, Plasmids and Primers
• cassette primers 2016
• Protocols
  • MIE review.pdf
  • Oligo optimization.pdf
  • Sharan-2009 Nature Protoc.pdf
  • high throughput.pdf
  • Thomason-2007 Plasmid.pdf
  • Thomason-2009 Met MB.pdf
  • preparing electrocompetent cells Lynno.pdf
  • transformation by electroporation.pdf
  • step-by-step ssDNA protocol.pdf
  • Brief ssDNA protocol.pdf
  • step-by-step knockout (dsDNA) protocol.pdf
  • brief dsDNA.pdf
  • checklist for plasmid recombineering.pdf
  • Thomason-2007 P1 prot.pdf
  • CPMB 2014.pdf
• References
  • General Recombineering
    • Datta-2006 Gene.pdf
    • Datta-2008 PNAS .pdf
    • Bubunenko-2007 J. bact.pdf
    • Yu-2003 PNAS.pdf
    • Court-2003 Gene.pdf
    • Yu-2000 PNAS.pdf
    • highthroughput.pdf
    • MIE 2013 dsDNA.pdf
    • CPMB 2014.pdf
    • LI Mol Micro 2013 with supp.pdf
  • High Throughput
  • ssDNA recombineering
  • Recombineering to BACs.
  • Recombineering to plasmids.
  • Recombineering to phage.
  • "Recombinase"-independent recombineering.
  • Recombineering reviews.
  • Oligo optimization.pdf
  • Li-2003 Nuc acid res.pdf
  • Costantino-2003 PNAS.pdf
  • Ellis-2001 PNAS.pdf
  • Thomason-2007 Plasmid.pdf
  • Thomason-2009 Met MB.pdf
  • Oppenheim-2004 Virol.pdf
  • Swingle 2010.pdf
  • Sharan-2009 Nature Protoc.pdf
  • MIE review.pdf
  • CurrentProtocols.pdf
  • Courtetal2002.pdf
  • Copeland-2001 Nat rev Genet.pdf
  • Warming-2005 Nuc acid res.pdf
  • Swaminathan-2001 Genesis.pdf
  • Lee-2001 Genomics.pdf
  • MIE 2013 ssDNA.pdf
  • CPMB 2014.pdf
  • Nucl. Acids Res.-2013-Li-e204
  • Fu et al. RecET
• Useful links
• Court Lab
• Photo 2
• Contact Form
• pSIM5.pdf
• pSIM6.pdf
• pSIM7.pdf
• pSIM8.pdf
• pSIM9.pdf
• pSIM10.pdf
• pSIM17.pdf
• pSIM18.pdf
• pSIM19.pdf
• pSIM27.pdf
• pSIM29.pdf
• tet-sacB map.pdf
• pSIM5.txt.zip
• pSIM6.txt.zip
• pSIM7.txt.zip
• pSIM8.txt.zip
• pSIM9.txt.zip
• pSIM10.txt.zip
• pSIM17.txt.zip
• pSIM18.txt.zip
• pSIM19.txt.zip
• pSIM27.txt.zip
• pSIM29.txt.zip
• tet-sacB.txt.zip
• pSIM5.zip
• pSIM6.zip
• pSIM7.zip
• pSIM8.zip
• pSIM9.zip
• pSIM10.zip
• pSIM17.zip
• pSIM18.zip
• pSIM19.zip
• pSIM27.gcc.zip
• pSIM29.gcc.zip
• tet-sacB.gcs.zip
• Recombineering Strains.docx
• 4-17-09 02.tif
• lab sans Don.JPG
• Hhs.Gov
• Nih.Gov
• Cancer.Gov
• Usa.Gov
• Ccr.Cancer.Gov