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Team:Cambridge/Lab book/Week 9
From 2012.igem.org
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===Tuesday (21/08/12)=== | ===Tuesday (21/08/12)=== | ||
| + | '''[[Team:Cambridge/Protocols/PCRProtocol|PCR of positive control]]''' | ||
| - | '''PCR for Mg Riboswitch''' | + | ---- |
| + | |||
| + | *Our order of Phusion has arrived- though we are still not confident with our PCR so we decided to first test with sfGFP | ||
| + | |||
| + | *Standard PCR conditions are used | ||
| + | |||
| + | *Results: success | ||
| + | |||
| + | |||
| + | |||
| + | '''Ribosense: PCR for Mg Riboswitch''' | ||
---- | ---- | ||
Revision as of 01:45, 27 September 2012
| Week: | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 13 | 14 |
|---|
Contents |
Monday (20/08/12)
Ratiometrica-Flu: 4th attempt at PCR of fragments
- Only vectors this time as we are still not sure if we have got the Velocity mastermix recipe right, as we are waiting for our orders of Phusion to arrive
- Results: no bands for any lanes including positive control
Tuesday (21/08/12)
- Our order of Phusion has arrived- though we are still not confident with our PCR so we decided to first test with sfGFP
- Standard PCR conditions are used
- Results: success
Ribosense: PCR for Mg Riboswitch
- PCRs were run for pJS130.1 split vector ('+8 codons')
- Colony PCR for endogenous Bacillus subtilis Mg Riboswitch ('+8 codons') was run
- Gels were run for above reactions, only one band for half of the pJS130 vector was visible.
Wednesday (22/08/12)
PCR for Mg Riboswitch
- PCRs were run for F Riboswitch from Yale's plasmid with overhangs for standard assembly.
- Colony PCR for endogenous Bacillus subtilis Mg Riboswitch (both '+8 codons' and '-8 codons' versions) were re-run.
- Gels were run for above reactions, only one band for positive control was visible.
Cells cultured for Miller assay
- B. subtilis strains carrying F riboswitch and 'KO' strains cultured separately in LB and 5µg/mL Chloramphenicol at 37°C
- E.Coli carrying F riboswitch cultured in LB and 50mg/ml Ampicillin at 37°C
Thursday (23/08/12)
- Miller assay set up and running with the wild type and crcB knock-out Bacillus strains and an E. coli strain which contains the construct on an episomal plasmid.
Friday (24/08/12)
