David Tulga Notebook
From 2007.igem.org
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Did the first cloning of the part Bca1145 from Bca1144. | Did the first cloning of the part Bca1145 from Bca1144. | ||
This involved first PCR'ing out the modified RFP gene from the p9145-Bca1144#5, then digestion and purification of the vector p9145. | This involved first PCR'ing out the modified RFP gene from the p9145-Bca1144#5, then digestion and purification of the vector p9145. | ||
| - | We then transformed some DH10 cells with the new part, and | + | |
| - | We then miniprep'd them, as well as did colony PCR. | + | We then transformed some DH10 cells with the new part, and cutured 2 clones. |
| + | We then miniprep'd them, as well as did digestion and colony PCR. | ||
Our colony PCR was unsuccessful, but the miniprep digestion indicated our part was correct. | Our colony PCR was unsuccessful, but the miniprep digestion indicated our part was correct. | ||
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We then sequenced the data and found that both sequencing files appear correct, although there was a possible sequencing error of calling an incorrect number of repeating AAA's in clone #1. | We then sequenced the data and found that both sequencing files appear correct, although there was a possible sequencing error of calling an incorrect number of repeating AAA's in clone #1. | ||
| + | |||
Sequencing Files: [[BerkiGEM2007-Sequencing-AY001 | AY001]], [[BerkiGEM2007-Sequencing-AY002 | AY002]] | Sequencing Files: [[BerkiGEM2007-Sequencing-AY001 | AY001]], [[BerkiGEM2007-Sequencing-AY002 | AY002]] | ||
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This includes the first few genes for the controller, including the low copy controller plasmid that will control the inducible BAC. | This includes the first few genes for the controller, including the low copy controller plasmid that will control the inducible BAC. | ||
| - | I also transformed some TG1 cells with the plasmid pSB4A3-I7101, for a | + | I also transformed some TG1 cells with the plasmid pSB4A3-I7101, for a mediprep of the low-copy plasmid we'll use for pI716101. I also cultured them in 100ml of Amp LB. |
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Revision as of 01:40, 6 June 2007
My Construction Files
My Sequencing Files
Dtulga - May 30th - June 5: Group Work
Did the first cloning of the part Bca1145 from Bca1144. This involved first PCR'ing out the modified RFP gene from the p9145-Bca1144#5, then digestion and purification of the vector p9145.
We then transformed some DH10 cells with the new part, and cutured 2 clones. We then miniprep'd them, as well as did digestion and colony PCR. Our colony PCR was unsuccessful, but the miniprep digestion indicated our part was correct.
We then sequenced the data and found that both sequencing files appear correct, although there was a possible sequencing error of calling an incorrect number of repeating AAA's in clone #1.
Sequencing Files: AY001, AY002
Dtulga - June 4-5: Independent Work
Designed construction files for pI716101 (Low Copy ampR Plasmid - Produces RFP), pir I716102 (R6K Origin Inducer), and T7rnap I716103 and I716104 (T7 RNA Polymerase).
This includes the first few genes for the controller, including the low copy controller plasmid that will control the inducible BAC.
I also transformed some TG1 cells with the plasmid pSB4A3-I7101, for a mediprep of the low-copy plasmid we'll use for pI716101. I also cultured them in 100ml of Amp LB.
to do
