IGEM:Harvard/2006/Cyanobacteria/Notebook/2006-10-13

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

1. • 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

1. • 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:

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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

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Notes: There is plenty of P3 left.

Also, midipreps of B18 and M2-7 are complete.

The nanodrop results are as follows: