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Team:BYU Provo/Notebook/CRISPR/febapr
From 2014.igem.org
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| + | <p>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. | ||
| + | <br/>- Using E coli DH5α we added in 1 µL of igem plasmid, incubated on ice for 2 min | ||
| + | <br/>- Heat shock at 42C for 60 sec. | ||
| + | <br/>- Recovered on Ice for 5 minutes | ||
| + | <br/>- Incubated for 30 minutes at 37C | ||
| + | <br/>- Plated 100 µL on a plate with chloramphenicol. 400 µL on another plate with chloramphenicol | ||
| + | <br/><ul> ○ 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. | ||
| + | <br/> ○ We will need to pick a colony and start an overnight from that to do plasmid preps from next class | ||
| + | </p> | ||
</html> | </html> | ||
Revision as of 21:30, 9 July 2014
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
