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Team:BYU Provo/Notebook/CRISPR/febapr
From 2014.igem.org
BYU 2014 Notebook |
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4/27/14 - Garrett Jensen.
Today we worked on amplifying our plasmid from the igem registry so that we have it ready to use when we get our crispr ready to move into e coli.
- Using E coli DH5α we added in 1 µL of igem plasmid, incubated on ice for 2 min
- Heat shock at 42C for 60 sec.
- Recovered on Ice for 5 minutes
- Incubated for 30 minutes at 37C
- Plated 100 µL on a plate with chloramphenicol. 400 µL on another plate with chloramphenicol
- ○ The plasmid is very concentrated, so transformation is usually very efficient. Plating all 500 µL of our bacteria would give us a whole lawn of bacteria, so we only will plate 100 so we can get individual colonies.
○ We will need to pick a colony and start an overnight from that to do plasmid preps from next class
