IGEM:MIT/2007/Notebook/2007-7-11: Difference between revisions
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:*Save some for standard assembly with sticky insert | :*Save some for standard assembly with sticky insert | ||
*Permanent glycerol stock (2ml for 2 stocks) | *Permanent glycerol stock (2ml for 2 stocks) | ||
===DD part E1010=== | |||
*Digested with EcoRI and SpeI | |||
*Used 121.5 ng/µL eppendorf of E1010 | |||
*Mixture: | |||
**8.23µL of DNA | |||
**5µL of EcoRI Buffer | |||
**2µL of EcoRI | |||
**2µL of SpeI | |||
**0.5µL of BSA | |||
**32.27µL of H2O | |||
*3 Hrs in 37C room. Put in at 11:50AM | |||
===Made More 1% Agarose in TAE Buffer=== | ===Made More 1% Agarose in TAE Buffer=== | ||
Line 22: | Line 34: | ||
*Agarose powder can be found on chemical powder shelf rm 558e. Containers are alphabetized. | *Agarose powder can be found on chemical powder shelf rm 558e. Containers are alphabetized. | ||
*Flask kept in the 55C box next to Jason's bay. | *Flask kept in the 55C box next to Jason's bay. | ||
===Results for Transformed XL-1 (pSB3K3 + F2620 + B0034) Miniprep=== |
Revision as of 12:44, 11 July 2007
Agenda
- 1.) Miniprep Backbone LC, DD eRFP (+, - controls), DD miniprep backbone (do we need to gel or pcr purify?), Standard assembly by running gel, purifying gel, ligation, transformation
- 2.) Make LC of MerPR and MerR Plates
- 3.) Send final transformation miniprep for sequencing
Diluted LC's of Transformed XL-1
- Used Samples #3 and #4 from overnight LC's
- 1:5 dilution: therefore, we now have 25µl for each cell line
- JH: Accidentally diluted line #3 6X (30µL total). Line #4 is still 5X dilution.
- Placed in 37C around 10am
Notes on Future Use of Transformed XL-1
- Miniprep
- Some for sequencing
- standard assembly with eRFP
- Save some for standard assembly with sticky insert
- Permanent glycerol stock (2ml for 2 stocks)
DD part E1010
- Digested with EcoRI and SpeI
- Used 121.5 ng/µL eppendorf of E1010
- Mixture:
- 8.23µL of DNA
- 5µL of EcoRI Buffer
- 2µL of EcoRI
- 2µL of SpeI
- 0.5µL of BSA
- 32.27µL of H2O
- 3 Hrs in 37C room. Put in at 11:50AM
Made More 1% Agarose in TAE Buffer
- 1L flask contains: 1L of TAE Buffer and 10 mg of agarose powder each
- TAE buffer can be found above the sink
- Agarose powder can be found on chemical powder shelf rm 558e. Containers are alphabetized.
- Flask kept in the 55C box next to Jason's bay.