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Team:Paris Bettencourt/Notebook/Phage Sensor/Wednesday 7th August.html
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Revision as of 10:28, 23 August 2013
Phage Sensor
ASDFWednesday 7th August
Glycerol Stock of sSP008-sSP0011, M13 Test, Single Colonies, Liquid cultures, PCR and Transformation
Glycerol Stock of sSP008-sSP0011Take 1.5ml of ON culture + 500ul Glycerol (60%)
Freeze at -80°C
M13 Test
Plate from ON culture:
1) 1:1000 - sSP012: sSP009 (plate 200ul)
2) 200 ul sSP012
3) 200ul sSp009
Single Colonies
streak out ON culture of sSP012 to get single colonies
Liquid cultures of pir+ E.coli and pKD13 from Jake
PCR of SPCR4, SPCR7, SPCR8, SPCR9, SPCR10 from circuar backbone, directly taken from Biobrick plate
Protocol
| Reagent | Volume |
| 1x | |
| Nuclease-free water | 37,25µL |
| 10µL | |
| 10 mM dNTPs | 1µL |
| Forward Primer (10 µM) | 0.5µL |
| Reverse Primer (10 µM) | 0.5µL |
| Template Plasmid | 0.25µL |
| Phusion DNA Polymerase | 0.5µL |
| Total Volume | 50µL |
