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Team:Paris Bettencourt/Notebook/Phage Sensor/Wednesday 7th August.html
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| - | <b>Glycerol Stock of sSP008-sSP0011</b><br> | + | <br><b>Glycerol Stock of sSP008-sSP0011</b><br> |
Take 1.5ml of ON culture + 500ul Glycerol (60%)<br> | Take 1.5ml of ON culture + 500ul Glycerol (60%)<br> | ||
Freeze at -80°C<br> | Freeze at -80°C<br> | ||
| Line 26: | Line 26: | ||
<b>Liquid cultures</b> of pir+ E.coli and pKD13 from Jake<br> | <b>Liquid cultures</b> of pir+ E.coli and pKD13 from Jake<br> | ||
<br> | <br> | ||
| - | <b>PCR of SPCR4, SPCR7, SPCR8, SPCR9, SPCR10 from circuar backbone, directly taken from Biobrick plate</b> | + | <b>PCR of SPCR4, SPCR7, SPCR8, SPCR9, SPCR10 from circuar backbone, directly taken from Biobrick plate</b><br><br> |
| - | <br> | + | |
Protocol<br> | Protocol<br> | ||
<br> | <br> | ||
| - | |||
<TABLE BORDER> | <TABLE BORDER> | ||
<TR><TD><b>Reagent</b></TD><TD><b>Volume</b></TD></TR> | <TR><TD><b>Reagent</b></TD><TD><b>Volume</b></TD></TR> | ||
<TR><TD> </TD><TD><b>1x</b></TD></TR> | <TR><TD> </TD><TD><b>1x</b></TD></TR> | ||
<TR><TD>Nuclease-free water</TD><TD>37,25µL</TD></TR> | <TR><TD>Nuclease-free water</TD><TD>37,25µL</TD></TR> | ||
| - | <TR><5x Phusion HF Buffer</TD><TD>10µL</TD></TR> | + | <TR><TD>5x Phusion HF Buffer</TD><TD>10µL</TD></TR> |
<TR><TD>10 mM dNTPs</TD><TD>1µL</TD></TR> | <TR><TD>10 mM dNTPs</TD><TD>1µL</TD></TR> | ||
<TR><TD>Forward Primer (10 µM)</TD><TD>0.5µL</TD></TR> | <TR><TD>Forward Primer (10 µM)</TD><TD>0.5µL</TD></TR> | ||
Revision as of 10:31, 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-sSP0011
Take 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 |
| 5x Phusion HF Buffer | 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 |
