Jessica Karen Wong/Notebook/2007-7-12

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