Revision as of 21:43, 15 August 2011

PROJECT

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.

@@ Line 6: / Line 6: @@
 <img src="https://static.igem.org/mediawiki/2011/4/40/Notebook_banner4_2.gif" height = "auto" width="750px" style="opacity:1;filter:alpha(opacity=100)" alt="NU-igem banner"/ border="0">
 <div style="margin: -55px 0px 0px 80px;font:35px helvetica; color:#ffffff;">
-Notebook   <div style="margin: -40px 0px 0px 420px;font:35px helvetica; color:#444444;"> Week 9</div>
+Notebook   <div style="margin: -40px 0px 0px 420px;font:35px helvetica; color:#444444;">    &nbsp; Week 9</div>
 </div></div>
 </html>