Registry/Measurement kit/Notebook/2007-7-2

Key

 * I2055 - GFP RBS tester
 * I2056 - RFP RBS tester (blue)
 * I2057 - RFP promoter tester (green)

Retransformation of I2057

 * Used previous ligation (tet and chlor)
 * Plated 200uL

Sequence check of I2057

 * dnalims.mit.edu
 * Mix for sequencing rxn
 * 1uL DNA (200-500 ng)
 * 2uL 1.6 uM VF2/VR
 * 9uL H2O

Scarring PCR

 * Gradient PCR of I2055 from 41 to 51 deg
 * PCR of T9002 with Tm of 54.5 deg

Primer Design
To insert BB sites. Format: 8-mer.restriction site.primer


 * E0240:
 * F (incl E0240, PstI): CTTAGTAG.ACTAGT.TCACACAGGAAAGTACTAGATGCG (53.1)
 * R (incl 3K3/1AK3, EcoRI): TCAGCGAT.GAATTC.CAGAAATCATCCTTAGCGAAAGC (54.2)


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


 * I2055:
 * F (incl GFP, SpeI): CTTAGTAG.ACTAGT.ATGCGTAAAGGAGAAGAACTTTTC (52.4)
 * R (incl R0040, EcoRI): TCAGCGAT.GAATTC.GTGCTCAGTATCTCTATCACTGATAGG (52.0)


 * I2056:
 * F (incl RFP, SpeI): CTTAGTAG.ACTAGT.ATGGCTTCCTCCGAAGACG (54.0)
 * R: Same as I2055


 * For T9002:
 * F (incl lux pR, PstI): TCAGCGAT.CTGCAG.TCACACAGGAAAGTACTAGATGCG (53.1)
 * R (incl RBS, GFP,EcoRI) : CTTAGTAG.GAATTC.TTTATTCGACTATAACAAACCATTTTC (51.9)