Registry/Measurement kit/Notebook/2007-7-2: Difference between revisions
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==Sequence check of I2057== | ==Sequence check of I2057== | ||
*dnalims.mit.edu | *dnalims.mit.edu | ||
*Mix for sequencing rxn | |||
**1uL DNA (200-500 ng) | **1uL DNA (200-500 ng) | ||
**2uL 1.6 uM VF2/VR | **2uL 1.6 uM VF2/VR | ||
Revision as of 12:47, 2 July 2007
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:
- For E0240:
- F (incl E0240): tail (PstI) + TCACACAGGAAAGTACTAGATGCG
- R (incl 3K3): tail (EcoRI) + AATTCCAGAAATCATCCTTAGCG
- For I2055:
- F (incl GFP): tail (speI) + ATGCGTAAAGGAGAAGAACTTTTC
- R (incl R0040): tail (ecorI) + GTGCTCAGTATCTCTATCACTGATAGG
