Team:TUDelft/1 September 2009

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='''1 September 2009'''=
='''1 September 2009'''=
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===Sriram===
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Today I continued with the final assembly of Riboregulator. Since Daniel is leaving I also continued his assembly of locks and keys. Today I sent samples to sequenceing (Medium RBS control, Plasmid 1 of negative cascade, Plasmid 2 of negative cascade and Weak RBS Control). I added both primers given by TIm Weenink.
===Daniel===
===Daniel===
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| Part || [DNA] ng/uL  
| Part || [DNA] ng/uL  
|- align="center"
|- align="center"
-
| 1 ||
+
| 1C || 8.5
|- align="center"
|- align="center"
-
| 1 ||
+
| 2C || 45.6
|- align="center"
|- align="center"
-
| 1 ||
+
| 3C || 20.5
|- align="center"
|- align="center"
-
| 1 ||
+
| 4C || 16.6
 +
|- align="center"
 +
| 1A || 161.2
 +
|- align="center"
 +
| 1B || 84.9
 +
|- align="center"
 +
| 2B || 95.2
 +
|- align="center"
 +
| 3B || 265.1
 +
|- align="center"
 +
| CP || 96.4
 +
|- align="center"
 +
| RLacI || 58.3
|}
|}

Latest revision as of 22:50, 21 October 2009

Lab Notebook

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1 September 2009

Sriram

Today I continued with the final assembly of Riboregulator. Since Daniel is leaving I also continued his assembly of locks and keys. Today I sent samples to sequenceing (Medium RBS control, Plasmid 1 of negative cascade, Plasmid 2 of negative cascade and Weak RBS Control). I added both primers given by TIm Weenink.

Daniel

Miniprep of yesterday's cultures:

Part [DNA] ng/uL
1C 8.5
2C 45.6
3C 20.5
4C 16.6
1A 161.2
1B 84.9
2B 95.2
3B 265.1
CP 96.4
RLacI 58.3

As noticed, 1C concentration is low, this is because I add neutralization buffer before lysis buffer. We will do it again tomorrow.Thanks to Esengul, we will have electrocompetent cells which produce LacI.

Today I leave the lab and I'll be helping in planning and experimental design.

Tim Weenink

Today I restricted p(LacI) as an upstream part and *T1 (homing endonuclease without promotor) as a downstream part. Additionally I ligated the restricted I-SceI PCR fragment. This fragment was PCR'd from the plasmid from leiden containing the conding sequence of ISCEI. It was purified and restricted using E and P. Using the standard protocol, it was ligated into a pSB1A3 vector.

Finally, Esengul streaked a plate of K12 strain, that allows IPTG induction (without being expressed constitutively). This strain wil be grown up and made electro competent.