User:Hetmann/Notebook/2006-6-20

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Performed a miniprep on the R0010 + E0241 inoculations from 6/19.

Created glycerol stocks of two cultures.

Creating Glycerol Stocks Protocol

• Obtain a screwcap-top eppendorf tube.
• Add 100 µL 50% glycerol.
• Add 100 µL liquid culture.

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When the 2 colonies were centrifuged the pellets fluoresced green.

Made minipreps of the two pelleted cultures using the Qiagen protocol.

Performed digest on the R0010 + E0241 using XbaI and PstI

Digestion Protocol

• Create 4 µL stock of 1 µL Xba + 1 µL Pst1 + 2 µL dH20 (Restriction Enzyme Stock)
• Create 4 µL stock of 1 µL BSA + 3 µL dH20 (BSA Stock)

• Pipet the following into the two tubes:
  • 8 µL DNA
  • 2 µL Restriction Enzyme Stock
  • 1 µL BSA Stock
  • 2.5 µL buffer 3
  • 11.5 µL dH20

Total volume: 25 µL

• Incubate at 37 Celcius for 1 hour

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Jeff took care of the digestions after incubation: I left the lab at 12:30 PM.

I went on a trip to Torrance, CA, to visit my sister's graduation.