Team:UIUC-Illinois/Notebook/LabNotebook

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5/29/2012	- Designed 9 primers for LacI. LacZ. and eYFP. Prepared LB. - Inoculation of B0014.J23100.wtPUF. B0030.Q04121 - Transformed the psB1C3 biobrick into DH5a cells
5/31/2012	- Transformed C0012, I732005, E0030, I732017. Also transformed PSB1A3, PSB1AK3, PSB1K3. - Inoculated LB media with a psB1C3 colony from the plate grown the day before. - Met with Prof. Bhalerao to talk about PUF application and project direction (RNA scaffolding).
6/1/2012	- Short-term goal changed: make one testing construct, [rbs-puf-eYFP] cloned into P.protet plasmid with a ptet promoter and terminator - Began inoculation of C0012, I732005, E0030, I732017, PSB1A3, PSB1AK3, PSB1K3. Was incubated for 9 hours, then put in 4 degree room - Inoculated again due to the round-bottom tube being misplaced. Mini-prepped the inoculation.
6/2/2012	- Cultured [PUF+PIN], E0030, and P. protet. designed check primers for [rbs-puf-eYFP]
6/3/2012	- Inoculated with a psB1C3 colony in order to prepare a glycerol stock of the strain.

6/4/2012	- Threw out old, incomplete inoculations from Friday. - Transformed I732005, R0010. - Miniprepped Venus and E0030 and put gDNA in the -20 temp box. - Did new inoculations on C0012, E0030, I732017, PSB1A3, PSB1AK3, PSB1K3. - Made a glycerol stock of the psB1C3 strain.
6/5/2012	- Made glycerol stocks of C0012, E0030, I732017, PSB1A3, PSB1AK3, PSB1K3 and put them in the -80 freezer. - Transformation of I732005 failed, so replated with rest of culture. - Transformed J04500 and PSB3T5. - Minipreped PUF Amp and protet CM20 (spilled 1 tube of this) and put gDNA in the -20 degree in the temp box. - Inoculated R0010.
6/7/2012	- Set goal to create RNA scaffold by 6/15, begun design.

5/29/2012	- Dr. Ho will ship vectors from Korea on Friday. Should receive them Monday 6/4/12. - Found and read literature on why C4H doesn't express well in E.coli as well as investigating a protocol for expressing the BM3s in E.coli. - Prof Koffas will ship us pUCo-VvSTS-At4CL as one piece if the MTA goes through. - Mark Langston of Octochem has offered to buy some trans-resveratrol for us. p-Courmaric acid is approx $60/25g at TCI America.
5/31/2012	- Group reading over literature - Revised BM3 primer design - Miniprepped 5 cultures: B0030, B0014, Q04121, J23100, WT PUF and put gDNA in -20 temp box
6/1/2012	- Group reading over literature. - Revised BM3 primer design again.
6/2/2012	- Literature review
6/3/2012	- Literature review

6/4/2012	- Received BM3 vectors from Prof. Ho as dried samples. - Submitted primer order to Courtney for ordering from IDT. - Literature review. - Planning to extract vector from well-plate.
6/5/2012	- Ordered the part BBa_K122012 from the parts registry, this is 4CL:STS with yeast promoter and terminator. - Informed by the Kim paper, the BM3s are under the control of an IPTG promoter. - Talked to Dr. Rao about making a protocol for testing the BM3s once they are in an expression vector, Dr. Rao wants PHAT group to lyse the cells and try an in vitro assay on the BM3 enzymes first. - Also found the ATCC company sells Rhodobacter capsulatus gDNA which could be useful for PCR of TAL gene. Parts registry does not have a TAL gene for us to order.
6/6/2012	- Read literature about TLC plates. Learned that we can get the plates from the chem store room.
6/7/2012	- Received feedback from Dr. Jin at the advisors meeting. - Will investigate whether TLC plates can separate piceatannol from resveratrol.

Revision as of 20:19, 13 July 2012