Jessica Karen Wong/Notebook/2007-7-12

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Sequencing

  • Minipreped overnight cultures of E0240-3K3, I2057-3K3, T9002-3K3, T9002-1AK3 for sequencing
  • Sequencing I2057-3K3 w/ VF and VR
    • E0240-3K3 with E0040F and E0040R
    • T9002-3K3 with E0040F and R
    • T9002-1AK3 with F2620F and R (aka c62VF and c62VR)
  • Made overnights of E0240-1AK3 and T9002-3K3 (to look for F2620) to sequence tomorrow

PCRs

  • Ran gel of overnight I2055 col PCR and E0240 BB pcr
  • Nothing was right
  • Redid E0240 BB PCR with Vent and a 3x dilution of DNA


Ordered New Primers

T9002

  • F2620-F 8mer mfe1 TCCCTATCAGTGATAGAGATTGACATC 53.9
  • CTTAGTAG CAATTG TCCCTATCAGTGATAGAGATTGACATC
  • F2620-R 8mer xba1 not1 eco1 TTTATTCGACTATAACAAACCATTTTC 51.9
  • TCAGCGAT CTCTAGAAGCGGCCGCGAATTC TTTATTCGACTATAACAAACCATTTTC


  • E0240-F 8mer spe1 NOT1 pst1 TCACACAGGAAAGTACTAGATGCG 53.1
  • CTTAGTAG tactagtagcggccgctgcag TCACACAGGAAAGTACTAGATGCG
  • E0240-R 8mer nsi1 GTGGGCCTTTCTGCGTTTATA 53.6
  • TCAGCGAT ATGCAT GTGGGCCTTTCTGCGTTTATA

I2055/I2056

  • I2055-F 8mer Xba1 ATGCGTAAAGGAGAAGAACTTTTC 52.4
  • CTTAGTAG TCTAGA ATGCGTAAAGGAGAAGAACTTTTC
  • I2055-R 8mer xba1 NOT1 ecor1 GTGCTCAGTATCTCTATCACTGATAGG 52.0
  • TCAGCGAT CTCTAGAAGCGGCCGCGAATTC GTGCTCAGTATCTCTATCACTGATAGG
  • I2056-F 8mer Xba1 ATGGCTTCCTCCGAAGACG 54.0
  • CTTAGTAG TCTAGA ATGGCTTCCTCCGAAGACG
  • I2056-R same as I2055-R