"
Team:Technion/26 August 2012
From 2012.igem.org
- Home
- Team
- Project
- Parts
- Notebook
- Safety
- Attributions & Achievements
- Human Practices
- Sponsors & Thanks
|
|
|
|
|
|
|
|
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Ilya
- Starters of tac+RBS+mCh in pSB1C3 and in pSB3C5, R0062+RBS+mCh in pSB1C3 and in pSB3C5 for PacI restriction test tomorrow.
- Found a biobrick with the T3 RNA polymerase, BBa_K346000. Apparently the registry search engine is not very efficient. I stumbled on the biobrick when looking through the inserts available in pSB1C3. Later, I tried looking for "T3 RNA polymerase site:partsregistry.org" in google and the biobrick was the first search result. Next time you look for biobricks, use google.
Inbal
- Ordered the new primers for T7 RNAP from chris. Also, I designed the final primers for the Gibson Assembly- hope that it will turn out good, Now I'm waiting for the results from the SP6 sequencing, before I will order the primers for Gibson. :)
- I digested the pTetO+GFP+ter (BBa_I13522) with EcoRI and PstI for 15 minutes at 37C, ran the products on 1% agarose gel for 30 minutes, The expected results were: 2 bands- one is 2000bp and the second is 940 bp approximately.
The actual results were completely not logical- I got bands at 1500bp and 3000bp, Also in the control lane there were number of bands (The control was the uncut plasmid). My conclusion for the this past two weeks, while working with BBa_I13522 is that something wrong with it. Tomorrow I will start working with a different BioBrick, one that contains pTetO and a reporting gene.
Asaf
I prepared starters of the different plasmids that contain the polymerase genes again (T7*,T3,K1F,N4,SP6).
Hila
Phage project:
PCR reaction for the phage lambda fragments that we plan to create.
The reaction preformed in the gradient PCR with the following annealing temperatures:
Fragment 1+1c – 63.2C
Fragment 2+2c – 63.7C
Fragment 3+3c – 61.9C
Fragment 4+4c – 61.2C
Fragment 5+5c – 63.7C
Fragment 6+6c – 61.9C
Fragment 7+7c – 58.5C
Fragment 8+8c – 62.7C
The gel shows that fragment 3, 3C, 4, 4C, 5, 5C, 7, 7C, 8 worked.
pSB1C3+MCS project:
Starter for positive colonies 1 and 3 (with 344bp band).
Lior
PCR purification of cutting products (pT3 pN4 pK1F pSp6 and xyIE).
Ligation of p- plasmids with inserts + control: without insert for all plasmids.
Starters for done pT7 - inserts for sequencing and glycerol stock.
Noa
Evgeni
- Morning: I've chosen six colonies from the plate with pPROLar+Bba_015 transformed bacteria and made starters.
- Evnening: Using morning starters for making miniprep and glycerol stock of the transformed bacteria.
