From 2012.igem.org
Team:Technion/15_August_2012
Ilya
Inbal
Today I did mini-preps for the starters from yesterday (TOP10 bacteria cells including plasmid pSB1C3 with SP6 RNAP gene+terminator).
I stored the sample at -20C.
Also I forgot to set aside a portion for the glycerol stock- so I did all over again a starter of the cells mentioned above.
A while back- Hila did a glycerol stock of cells with the pSB1A2+pTetO+GFP+ 2 terminators (BBa_I13522), so today I plated those cells on LB+Amp plate and stored the plate in 37C incubator.
I helped Hila with the mini-prep of Fus1 and Fus2 (she helped to Ilya)
Also I started reading on the Gibson assembly- for fusing the pTetO+GFP to the SP6 gene+terminator. Tomorrow I'm going to plan the primers and order them ASAP.
Asaf and Rachel
We minipreped the plasmids that contain the different polymerases genes and promoters we received from Chris.
Hila
Restriction reaction using PstI and EcoRI for the parts:
1. pSB1A2+R0062_Fus1 <8a+8b> (2.5ug)
2. pSB3C5 <7b> (3ug)
3. pSB1C3+Fus2 <P1> (3ug)
Gel purification to the restriction products:
1. pSB1A2+R0062_Fus1: insert only (877bp) [8a – 13.1ng/ul; 8b – 13.3ng/ug]
2. pSB3C5: insert (859bp) + back bone (2029bp) [insert – 7.6ng/ul]
3. pSB1C3+Fus2: (2697bp)
Lior
Noa
Restriction for PCR products of ALP gene.
PCR purification for cut products.
Because concentration wasn't high enough, an additional PCR is needed for ALP and xyIE.
Evgeni
Shahar
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