Jessica Karen Wong/Notebook/2007-7-18

Biobrick Primers

• E0240:
  • F_New (incl E0240, Spe1): CTTAGTAG.ACTAGT.TCACACAGGAAAGTACTAGATGCG (53.1)
  • R Same as before (incl 3K3/1AK3, EcoRI): TCAGCGAT.GAATTC.CAGAAATCATCCTTAGCGAAAGC (54.2)

• I2057:
  • F (incl I2057, Spe1): CTTAGTAG.ACTAGT.TCACACAGGAAAGTACTAGATGGC (52.1)
  • R: same as E0240

T9002 E0240 piece

• E0240-F 8mer spe1 TCACACAGGAAAGTACTAGATGCG 53.1
• Same as E0240_BB_F

PCR/Gel

• Ran a gel of E0240-3K3 BB PCR and E0240_T9002 tails PCR - nothing showed up
• Made a Colony PCR of the 1 colony of F2620-3K3
  • Ran a gel of it and looks good - minipreped for sequencing
• PCR'ed another 100ul E0240_T9002 tails PCR at 51 with vent and a 1:30 ext time

Sequencing

• Got some of the redone sequencing from monday
• E0240-3K3 looks scarred on both sides
• I2055-3K3 is scared but has a random 1 base insertion towards the end
• I2055-1AK3 still has no promoter
• Minipreped I2055-3K3 (that we're pretty sure contains the promoter) for seq
• Shouldn't put too much faith in these b/c these results still aren't the ones we made glycerols of