Team:Technion/27 August 2012

From 2012.igem.org



Team:Technion/27_August_2012
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November
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Ilya

- Minipreped tac+RBS+mCh in pSB1C3 and in pSB3C5, R0062+RBS+mCh in pSB1C3 and in pSB3C5 (4 colonies each).
- Digested the above samples with PacI. There wasn't enough PacI so I used a low concentration of the enzyme.
- Ran the restriction products on a gel. Most of the samples weren't completely digested but it appears as if I have at least one good clone from each set.
- Prepared one clone from each set for sequencing.
- Prepared BA media for the plate reader on Wednesday.

Inbal

- Made 2 starters of BBA_I13600 (pTetO+GFP)from the glycerol stock Hila did few weeks ago. In the evening, I mini preped them and the concentrations were: 85 and 118 ng/ul.
-Also, I diluted the 7I well in the distribution kit number 4, which contains the BioBrick BBa_K346000. Tomorrow I will do transformation to the TOP10 bacteria.
-The results from SP6 sequencing came, only 1 out of 4 primers came with confirmed sequence- But the sequence wasn't compatible to the SP6 gene, but to pSB1C3 plasmid (the sequencing began at the vector).

Asaf

I minipreped the plasmids that contain the different polymerase genes
and checked their concentration (it wasn't any different from the previous one).

Hila

Phage project:
PCR reaction for fragments 1, 1C, 2, 2C, 6, 6C, 8C all in T annealing = 64C.
Fragments 2, 2C and 8C worked.

pSB1C3+MCS project:
Glycerol stock and mini-prep for colonies 1 and 3.
The mini-prep product will be sent tomorrow for sequencing.

Lior

We minipreped from starters and made glycerol stock.
Making CM plates for tranformation.

Noa

Evgeni

- Restriction analysis of the miniprep: Ligation of Bba_015 into the pPROLar backbone adds unique NheI restriction site.
- After running digested samples I've got no digestion at all in all 6 minipreps! (Sad face). Either there was no insert in the plasmids or there was some mistake in preparation of miniprep or restriction analyses.
- I'm trying to make new starters from bacteria in the glycerol stock in hope that absence of digestion was due to some mistake in miniprep and not due to self ligation.

Shahar

Minipreped tac+RBS+mCh in pSB1C3 and in pSB3C5, R0062+RBS+mCh in pSB1C3 and in pSB3C5 (4 colonies each).

Rachel