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Purdue/11 July 2008
From 2008.igem.org
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===Glycerol Stocks!=== | ===Glycerol Stocks!=== | ||
*Today making glycerol stocks of SOS (J22106) and LacZ (I732017) parts from stabs sent by iGEM | *Today making glycerol stocks of SOS (J22106) and LacZ (I732017) parts from stabs sent by iGEM | ||
| + | *We only have 100% glycerol (sterile), so we're just going to follow the same iGEM protocol but with 100% glycerol | ||
| + | *Dr. Clase also suggested a somewhat different protocol for making stocks, so we're going to do both side-by-side and see which is better | ||
*Following iGEM protocol: | *Following iGEM protocol: | ||
**Make o/n culture in antibiotic (amp) LB broth --did this yesterday | **Make o/n culture in antibiotic (amp) LB broth --did this yesterday | ||
| - | **Combine 1mL of overnight w/ 150 uL of sterile | + | **Combine 1mL of overnight w/ 150 uL of sterile 100% glycerol solution in screw-top cryotube (make sure tube is labeled with part #, plasmid, and antibiotic resistance) |
**Vortex briefly | **Vortex briefly | ||
**Incubate at room temp. for 1/2 hour and put in -80C freezer | **Incubate at room temp. for 1/2 hour and put in -80C freezer | ||
| + | *Following Dr. Clase's protocol: | ||
| + | *Mix 750uL 100% glycerol and 250uL stock colony in cryotube | ||
| + | *Vortex | ||
| + | *Put directly in -80C freezer | ||
| + | |||
In other news, the bead plating did not help the other Lac transformations to work, although the pUC19 still grew quite well. | In other news, the bead plating did not help the other Lac transformations to work, although the pUC19 still grew quite well. | ||
| + | |||
'''Edited by Janie Stine''' | '''Edited by Janie Stine''' | ||
Latest revision as of 17:31, 11 July 2008
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Glycerol Stocks!
- Today making glycerol stocks of SOS (J22106) and LacZ (I732017) parts from stabs sent by iGEM
- We only have 100% glycerol (sterile), so we're just going to follow the same iGEM protocol but with 100% glycerol
- Dr. Clase also suggested a somewhat different protocol for making stocks, so we're going to do both side-by-side and see which is better
- Following iGEM protocol:
- Make o/n culture in antibiotic (amp) LB broth --did this yesterday
- Combine 1mL of overnight w/ 150 uL of sterile 100% glycerol solution in screw-top cryotube (make sure tube is labeled with part #, plasmid, and antibiotic resistance)
- Vortex briefly
- Incubate at room temp. for 1/2 hour and put in -80C freezer
- Following Dr. Clase's protocol:
- Mix 750uL 100% glycerol and 250uL stock colony in cryotube
- Vortex
- Put directly in -80C freezer
In other news, the bead plating did not help the other Lac transformations to work, although the pUC19 still grew quite well.
Edited by Janie Stine
