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Team:Northwestern/Notebook/Week9
From 2011.igem.org
PROJECT
RESULTS
CONSIDERATIONS
ABOUT US
NOTEBOOK
ATTRIBUTIONS
Notebook
Week 9
Day 39 - Monday, August 8th 2011
- Miniprepped overnight cultures from the attempted Oligo based ligations last week.
- Digested the minipreps and ran them on a gel to look for the correct sized bands.
- Prepared samples for sequencing
- Made new Tet Plates, Chlor Plates and SOB Broth
- PCRed our strep tag primer onto our CP->LasR and CP-> RhlR constructs
Day 40 - Tuesday, August 9th 2011
- Made a new batch of competent cells. We tried to get them in the -80 freezer extremely quickly to improve the efficiency.
- We digested our strep tag PCR products with DpnI and then added ligase so the blunt ends would come back together. We ligated one batch for one hour and another overnight.
- We transformed the overnight ligations
- We transformed to test the competency of our new competent cells
- Sent in samples for seqencing
- Designed our T-shirts!
- Rafay gave a presentation on the math model
Day 41 - Wednesday, August 10th 2011
- Our competent cell test had only 21 colonies, indicating a very low efficiency =(
- Our transformations of the strep tag ligations also failed, although we can still try transforming the overnight ligations.
- Hopefully we’ll get sequencing results soon....
- Miniprepped the overnights from our glycerol stocks and of our terminators
- Transformed the strep tag overnight ligations from yesterday
Day 42 - Thursday, August 11th 2011
- The overnight ligations of the strep tag constructs produced four colonies! These were overnighted for miniprep tomorrow.
- Ran another digest of the CP -> RBS -> LasR/RhlR -> Strep tag constructs, this time after phosphorylating the ends of the DNA. This should help improve ligation efficiency.
- Ligated those digestions overnight
- Ran our strep tag PCR products on a gel to try and confirm that the initial amplification was successful.
- Prepared overnights of RBS+Part ligations from glycerol stocks and transformation plates. We will ligate our genomic promoters to these tomorrow.
- Started 3 test ligations with Genomic Promoter -> RBS -> Part from already existing stocks.
- Made a lot of progressing designing our human practices project
- The team and sponsorship pages on the wiki are nearing completion.
Day 43 - Friday, August 12th 2011
- Transformed the strep tag ligations
- Miniprepped overnights of RBS34+Part Ligations
- We got our sequencing results back! Most of the samples look very good and closely match what we expected.
- Made some more progress on the website.
