From 2014.igem.org

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Revision as of 14:12, 15 October 2014

Cornell iGEM

Notebook

Week 1 (June 2 - June 8)

Wetlab Overview

Transformation efficiency and protocols were tested - heat shock did not work well, and we came to the conclusion to stick with electroporation. The first day of wetlab training began on the 7th! We started working on amplification of metallothionein genes - so far nixA and GST-PMT appear to be working.
Drylab Overview

Text goes here.

Week 2 (June 9 - June 15)

Wetlab Overview

Obtained transformants of pA14F and pC14T heat shocks and both E/B and E/D were successfully heat shocked into DH5a. Ran into problems later in the week with heat shocked plates from overgrowth and contamination; switched to electroporating of C+F and E+B.

Lead Subteam

Text

Mercury Subteam

Text

Nickel Subteam

Text

Reporters Subteam

Text
Drylab Overview

Text goes here.

Week 3 (June 16 - June 22)

Wetlab Overview

This week we ligated various parts (F+A, F+C, E+B, and E+D), transformed, ran colony PCRs, and submitted them for sequencing. Most of our attempts were unsuccessful either due to failed ligations or contamination. We also made T for future minipreps.

Lead Subteam

Text

Mercury Subteam

Text

Nickel Subteam

Text

Reporters Subteam

Text
Drylab Overview

Text goes here.

Week 4 (June 23 - June 29)

Wetlab Overview

Cloning and troubleshooting continues. E+D, F+C, E+D, E+B4, and E+D11 sequencing results came in negative. The restriction cocktail method was used on F+C and F+A because mRFP kept religating back into the backbone. Plates were successful, but colony PCRs showed they were negative. Repeat cloning. We are facing problems with contamination of lead transporter and R plate. Submitting F+C3 for sequencing. We are also working on transforming off the AA and AB kit plate.

Lead Subteam

Text

Mercury Subteam

Text

Nickel Subteam

Text

Reporters Subteam

Text
Drylab Overview

Text goes here.

Week 5 (June 30 - July 6)

Wetlab Overview

Transformants gave negative signals; many gels lacked proper bands in correct locations. Growth assay looked good for mercury although concentration needs to be increased for nickel.

Lead Subteam

Text

Mercury Subteam

Text

Nickel Subteam

Text

Reporters Subteam

Text
Drylab Overview

Text goes here.

Week 6 (July 7 - July 13)

Wetlab Overview

We gathered the data for an assay testing the growth of cells in different heavy metals. In addition, we ligated each the lead transporter (R), and nickel (AA) and mercury (AB) inducible promoters with H. Unfortunately, our transformations did not seem to work for reasons like incomplete digests and inefficient stocks.

Lead Subteam

Text

Mercury Subteam

Text

Nickel Subteam

Text

Reporters Subteam

Text
Drylab Overview

Text goes here.

Week 7 (July 14 - July 20)

Wetlab Overview

We are troubleshooting some problems with gel purification and smearing on the gel when we visualize it. Our new stocks of competent cells also appear to be bad. Sequencing of R+H returned a portion of a random promoter region of Enterococcus, so this will have to be remade. We are in the process of creating many different BioBrick parts, such as AB1+H, F, AC+H, AB2+H, AC1S+H, AB2S+H, AC+H, AA + H, which are all at various stages of the cloning process.

Lead Subteam

Text

Mercury Subteam

Text

Nickel Subteam

Text

Reporters Subteam

Text
Drylab Overview

Text goes here.

Week 8 (July 21 - July 27)

Wetlab Overview

Text

Lead Subteam

Text

Mercury Subteam

Text

Nickel Subteam

Text

Reporters Subteam

Text
Drylab Overview

Text goes here.

Week 9 (July 28 - August 3)

Wetlab Overview

Text

Lead Subteam

Text

Mercury Subteam

Text

Nickel Subteam

Text

Reporters Subteam

Text
Drylab Overview

Text goes here.

Week 10 (August 4 - August 10)

Wetlab Overview

Text

Lead Subteam

Text

Mercury Subteam

Text

Nickel Subteam

Text

Reporters Subteam

Text
Drylab Overview

Text goes here.

Week 11 (August 11 - 17)

Wetlab Overview

Text

Lead Subteam

Text

Mercury Subteam

Text

Nickel Subteam

Text

Reporters Subteam

Text
Drylab Overview

Text goes here.

Week 12 (August 18 - August 24)

Wetlab Overview

Text

Lead Subteam

Text

Mercury Subteam

Text

Nickel Subteam

Text

Reporters Subteam

Text
Drylab Overview

Text goes here.

Week 13 (August 25 - August 31)

Wetlab Overview

Text

Lead Subteam

Text

Mercury Subteam

Text

Nickel Subteam

Text

Reporters Subteam

Text
Drylab Overview

Text goes here.

Week 14 (September 1 - September 7)

Wetlab Overview

Text

Lead Subteam

Text

Mercury Subteam

Text

Nickel Subteam

Text

Reporters Subteam

Text
Drylab Overview

Text goes here.

Week 15 (September 8 - September 14)

Wetlab Overview

Text

Lead Subteam

Text

Mercury Subteam

Text

Nickel Subteam

Text

Reporters Subteam

Text
Drylab Overview

Text goes here.

Week 16 (September 15 - September 21)

Wetlab Overview

Text

Lead Subteam

Text

Mercury Subteam

Text

Nickel Subteam

Text

Reporters Subteam

Text
Drylab Overview

Text goes here.

Week 17 (September 22 - September 28)

Wetlab Overview

Text

Lead Subteam

Text

Mercury Subteam

Text

Nickel Subteam

Text

Reporters Subteam

Text
Drylab Overview

Text goes here.

Week 18 (September 29 - October 5)

Wetlab Overview

Text

Lead Subteam

Text

Mercury Subteam

Text

Nickel Subteam

Text

Reporters Subteam

Text
Drylab Overview

Text goes here.

Week 19 (October 6 - October 12)

Wetlab Overview

Text

Lead Subteam

Text

Mercury Subteam

Text

Nickel Subteam

Text

Reporters Subteam

Text
Drylab Overview

Text goes here.

Week 20 (October 13 - October 19)

Wetlab Overview

Text

Lead Subteam

Text

Mercury Subteam

Text

Nickel Subteam

Text

Reporters Subteam

Text
Drylab Overview

Text goes here.

@@ Line 161: / Line 161: @@
 							<h4 class="media heading">Wetlab Overview</h4>
 							<p>
-								Text
+								Obtained transformants of pA14F and pC14T heat shocks and both E/B and E/D were successfully heat shocked into DH5a.  Ran into problems later in the week with heat shocked plates from overgrowth and contamination; switched to electroporating of C+F and E+B.
 							</p>
@@ Line 226: / Line 226: @@
 							<h4 class="media heading">Wetlab Overview</h4>
 							<p>
-								Text
+								This week we ligated various parts (F+A, F+C, E+B, and E+D), transformed, ran colony PCRs, and submitted them for sequencing. Most of our attempts were unsuccessful either due to failed ligations or contamination. We also made T for future minipreps.
 							</p>
@@ Line 290: / Line 290: @@
 							<h4 class="media heading">Wetlab Overview</h4>
 							<p>
-								Text
+								Cloning and troubleshooting continues. E+D, F+C, E+D, E+B4, and E+D11 sequencing results came in negative. The restriction cocktail method was used on F+C and F+A because mRFP kept religating back into the backbone. Plates were successful, but colony PCRs showed they were negative. Repeat cloning. We are facing problems with contamination of lead transporter and R plate.
+Submitting F+C3 for sequencing. We are also working on transforming off the AA and AB kit plate.
 							</p>
@@ Line 354: / Line 356: @@
 							<h4 class="media heading">Wetlab Overview</h4>
 							<p>
-								Text
+								Transformants gave negative signals; many gels lacked proper bands in correct locations.  Growth assay looked good for mercury although concentration needs to be increased for nickel.
 							</p>
@@ Line 418: / Line 420: @@
 							<h4 class="media heading">Wetlab Overview</h4>
 							<p>
-								Text
+								We gathered the data for an assay testing the growth of cells in different heavy metals. In addition, we ligated each the lead transporter (R), and nickel (AA) and mercury (AB) inducible promoters with H. Unfortunately, our transformations did not seem to work for reasons like incomplete digests and inefficient stocks.
 							</p>
@@ Line 482: / Line 484: @@
 							<h4 class="media heading">Wetlab Overview</h4>
 							<p>
-								Text
+								We are troubleshooting some problems with gel purification and smearing on the gel when we visualize it.  Our new stocks of competent cells also appear to be bad. Sequencing of R+H  returned a portion of a random promoter region of Enterococcus, so this will have to be remade. We are in the process of creating many different BioBrick parts, such as AB1+H, F, AC+H,  AB2+H, AC1S+H, AB2S+H, AC+H, AA + H, which are all at various stages of the cloning process.
 							</p>

Team:Cornell/notebook

From 2014.igem.org

Revision as of 14:12, 15 October 2014

Notebook

Week 1 (June 2 - June 8)

Wetlab Overview

Drylab Overview

Week 2 (June 9 - June 15)

Wetlab Overview

Lead Subteam

Mercury Subteam

Nickel Subteam

Reporters Subteam

Drylab Overview

Week 3 (June 16 - June 22)

Wetlab Overview

Lead Subteam

Mercury Subteam

Nickel Subteam

Reporters Subteam

Drylab Overview

Week 4 (June 23 - June 29)

Wetlab Overview

Lead Subteam

Mercury Subteam

Nickel Subteam

Reporters Subteam

Drylab Overview

Week 5 (June 30 - July 6)

Wetlab Overview

Lead Subteam

Mercury Subteam

Nickel Subteam

Reporters Subteam

Drylab Overview

Week 6 (July 7 - July 13)

Wetlab Overview

Lead Subteam

Mercury Subteam

Nickel Subteam

Reporters Subteam

Drylab Overview

Week 7 (July 14 - July 20)

Wetlab Overview

Lead Subteam

Mercury Subteam

Nickel Subteam

Reporters Subteam

Drylab Overview

Week 8 (July 21 - July 27)

Wetlab Overview

Lead Subteam

Mercury Subteam

Nickel Subteam

Reporters Subteam

Drylab Overview

Week 9 (July 28 - August 3)

Wetlab Overview

Lead Subteam

Mercury Subteam

Nickel Subteam

Reporters Subteam

Drylab Overview

Week 10 (August 4 - August 10)

Wetlab Overview

Lead Subteam

Mercury Subteam

Nickel Subteam

Reporters Subteam

Drylab Overview

Week 11 (August 11 - 17)

Wetlab Overview

Lead Subteam

Mercury Subteam

Nickel Subteam

Reporters Subteam

Drylab Overview

Week 12 (August 18 - August 24)

Wetlab Overview

Lead Subteam