IGEM:Harvard/2006/Cyanobacteria/Notebook/2006-10-13: Difference between revisions
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*B18 KaiB (J36011) \SP: 8 samples | |||
*M2-7 KaiC (J36010) \XP: 8 samples | |||
Protocols: | |||
J36011 \SP: 8 samples | |||
#*7.6 uL DNA (66.3) | |||
#*1 µL 10x BSA (8.5) | |||
#*1 µL 10x buffer #2 (8.5) | |||
#*0.2 µL Spe1 (1.7) | |||
#*0.2 µL Pst1 (1.7) | |||
J36010 \XP: 8 samples | |||
#*7.6 uL DNA (8.5) | |||
#*1 µL 10x BSA (8.5) | |||
#*1 µL 10x buffer #3 (8.5) | |||
#*0.2 µL Xba1 (1.7) | |||
#*0.2 µL Pst1 (1.7) | |||
Digest Protocol: | |||
------------ | |||
1) 45C, X Hours S.t. it ends ~1030AM Saturday (no more than 12 though) | |||
2) 80C, 20 min | |||
3) 4C infinity | |||
===CIP THE VECTOR=== | |||
BEFORE running the gel, '''not the insert''' (1uL CIP, 1h @ 37) | |||
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-Peng | -Peng | ||
------- | |||
Notes: There is plenty of P3 left. | |||
Also, midipreps of B18 and M2-7 are complete. | |||
The nanodrop results are as follows: |
Latest revision as of 19:02, 13 October 2006
Midipreps
- M2-7 KaiC, 25uL
- B18 KaiB, 25uL
Digestion
- B18 KaiB (J36011) \SP: 8 samples
- M2-7 KaiC (J36010) \XP: 8 samples
Protocols:
J36011 \SP: 8 samples
- 7.6 uL DNA (66.3)
- 1 µL 10x BSA (8.5)
- 1 µL 10x buffer #2 (8.5)
- 0.2 µL Spe1 (1.7)
- 0.2 µL Pst1 (1.7)
J36010 \XP: 8 samples
- 7.6 uL DNA (8.5)
- 1 µL 10x BSA (8.5)
- 1 µL 10x buffer #3 (8.5)
- 0.2 µL Xba1 (1.7)
- 0.2 µL Pst1 (1.7)
Digest Protocol:
1) 45C, X Hours S.t. it ends ~1030AM Saturday (no more than 12 though) 2) 80C, 20 min 3) 4C infinity
CIP THE VECTOR
BEFORE running the gel, not the insert (1uL CIP, 1h @ 37)
Hey Hetmann,
Sorry I had to leave immediately at 1; just to let you know tho, I only got to step 1 – pelleting; the cells were done pelleting at 1258 ish, so hopefully Brownian motion hasn’t affected them that much :D The little tubes on top of the centrifuge are clean. As for neutralization buffer (P3), we don’t have any more, but supposedly N3 is the same… I will try to call and confirm.
-peng
Damnit, I just called qiagen and the neutralization buffers ARE NOT the same; one is used for the miniprep tubes and the other for the QIAquick kit, which have different compositions =( Let Alain know and he should be able to buy another P3…
-Peng
Notes: There is plenty of P3 left.
Also, midipreps of B18 and M2-7 are complete.
The nanodrop results are as follows: