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Team:University of Lethbridge/Notebook/Project2August
From 2008.igem.org
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====Nathan Puhl, Roxanne==== | ====Nathan Puhl, Roxanne==== | ||
Ran riboswitch (Aug. 1, 2008) on 3% agarose gel. Results are in the hard copy lab notebook. Looks like 76 bp band will extract. | Ran riboswitch (Aug. 1, 2008) on 3% agarose gel. Results are in the hard copy lab notebook. Looks like 76 bp band will extract. | ||
| + | |||
| + | ===August 7, 2008=== | ||
| + | ====Nathan Puhl, Roxanne==== | ||
| + | Riboswitch: | ||
| + | |||
| + | Set up PCR using purified riboswitch from Aug. 5, 2008 with platinum Taq (50 uL reaction). Made Master Mix for three reactions. | ||
| + | Master Mix: | ||
| + | -10x Buffer (no Mg2+): 15 uL | ||
| + | -10 mM dNTPs: 3 uL | ||
| + | -50 mM Mg2+: 4.5 uL | ||
| + | -10uM RF: 3 uL | ||
| + | -10uM RR: 3 uL | ||
| + | -Plat. poly: 0.6 iL | ||
| + | -H20: 120.9 uL | ||
| + | -template: 1 uL | ||
| + | |||
| + | Cycle conditions: | ||
| + | A. Initial denaturation: 94 C (2 min) | ||
| + | B. -Denaturation: 94 C (30 sec) | ||
| + | - Annealing: 55 C (30 sec) | ||
| + | -Extension: 72 C (30 sec) | ||
| + | -30 cycles | ||
| + | C. Final extension: 72 C (7 min) | ||
| + | |||
| + | ====Roxanne==== | ||
| + | -Ran the PCR Product on a 3% gel using only 1uL of DNA from the riboswitch. | ||
Revision as of 23:14, 3 September 2008
Back to The University of Lethbridge Main Notebook
Contents |
August 1, 2008
Nathan Puhl, Roxanne
Riboswitch 20 uL PCR. Set up 4 reactions (25 uL for each total volume). 1 uL or 1/100 pTopp and 1 uL or H1/100 PCR from July 28, 2008.
PCR conditions:
A. Initial denaturation: 98 C (3 min)
B. -Denaturation: 98 C (10 sec)
- Annealing: 55 C (30 sec)
-Extension: 72 C (15 sec)
-30 cycles
C. Final extension: 72 C (7 min)
August 2, 2008
Nathan Puhl, Roxanne
Ran riboswitch (Aug. 1, 2008) on 3% agarose gel. Results are in the hard copy lab notebook. Looks like 76 bp band will extract.
August 7, 2008
Nathan Puhl, Roxanne
Riboswitch:
Set up PCR using purified riboswitch from Aug. 5, 2008 with platinum Taq (50 uL reaction). Made Master Mix for three reactions. Master Mix:
-10x Buffer (no Mg2+): 15 uL -10 mM dNTPs: 3 uL -50 mM Mg2+: 4.5 uL -10uM RF: 3 uL -10uM RR: 3 uL -Plat. poly: 0.6 iL -H20: 120.9 uL -template: 1 uL
Cycle conditions:
A. Initial denaturation: 94 C (2 min)
B. -Denaturation: 94 C (30 sec)
- Annealing: 55 C (30 sec)
-Extension: 72 C (30 sec)
-30 cycles
C. Final extension: 72 C (7 min)
Roxanne
-Ran the PCR Product on a 3% gel using only 1uL of DNA from the riboswitch.
