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Purdue/16 October 2008
From 2008.igem.org
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Therefore, both UV-exposed and non-exposed bacteria were turning blue because they were on X-gal plates. | Therefore, both UV-exposed and non-exposed bacteria were turning blue because they were on X-gal plates. | ||
| + | |||
| + | We were able to try a different method of transformation: Dr. Clase provided us with some Bio-Rad Hb101 cells (which naturally take up plasmids). We are following the Bio-Rad protocol for transformation: | ||
| + | # Put 250uL transformation solution (CaCl2) into an eppendorf tube | ||
| + | # Put tube on ice | ||
| + | # Add colony from starter plate of Hb101 cells to tube | ||
| + | # Incubate on ice 10 min. | ||
| + | # Heat shock at 42C for 50 seconds | ||
| + | # QUICKLY move back to ice and incubate for 2 minutes | ||
| + | # Remove from ice, add 250uL LB broth, incubate at RT for 10 min. | ||
| + | # Tap to mix, plate 100uL on amp plate | ||
'''Edited by Janie Stine''' | '''Edited by Janie Stine''' | ||
Latest revision as of 15:05, 16 October 2008
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Performed another ligation (for protocol, see Sept. 25). It turns out that the competent cells we used before are lacZ+. :(
Therefore, both UV-exposed and non-exposed bacteria were turning blue because they were on X-gal plates.
We were able to try a different method of transformation: Dr. Clase provided us with some Bio-Rad Hb101 cells (which naturally take up plasmids). We are following the Bio-Rad protocol for transformation:
- Put 250uL transformation solution (CaCl2) into an eppendorf tube
- Put tube on ice
- Add colony from starter plate of Hb101 cells to tube
- Incubate on ice 10 min.
- Heat shock at 42C for 50 seconds
- QUICKLY move back to ice and incubate for 2 minutes
- Remove from ice, add 250uL LB broth, incubate at RT for 10 min.
- Tap to mix, plate 100uL on amp plate
Edited by Janie Stine
