Team:Wageningen UR/Notebook/Proj1/August

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== August - Synchronised Oscillatory System ==
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|colspan="7"|<span style="color:white;">[https://2011.igem.org/Team:Wageningen_UR/Notebook/Proj1/July Previous Month]</span>
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== August - Synchronized Oscillatory System ==
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[[File:Aug.png]]
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'''August 2'''
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Colony PCR to verify ligations.
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Also some transformations.
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'''August 3'''
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Verifying parts using GFP and making liquid cultures from PCR- and GFP-confirmed colonies.
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'''August 4'''
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We isolated the following assembled parts: F2621-I763020 & R0062-I763020 & F2621-K082014-F2621-I763020 & F2621-I0460-F2621-I763020 & F2621-I0460-K318511 & R0062-I0460-K182102-K082014 & K182102-K082014-R0062-I763020
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This day we discovered something horrible... Our sequencing shows that the F2621 part from the registry did not have the R0063 promoter in front of it so at this point we realized we had to redo a lot of our cloning...
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'''August 5'''
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We isolated the following assembled parts:  K182102-C0061-B0015 & F2621-I0460
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'''August 6'''
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We isolated the assembled part: K182102-K082014 &
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'''August 8'''
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Digestions, ligations, transformations.
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'''August 10'''
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Digestions, ligations, transformations.
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'''August 11'''
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Yesterday’s gel showed that the digestions were a bit crappy so we redid all of them and then ligated and transformed again.
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Colony PCR of the transformants from the 10th of August.
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'''August 12'''
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Miniprepped the following parts: R0063-F2621-I0460 & R0063-F2621-I763020 & F2621-E0422-F2621-K082014 & R0063-F2621-K082014 & K176028-I0460
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Digestions were started to then be ligated and used to transform E. coli.
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'''August 13'''
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PCR screen of the ligations from August 12th.
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Also minipreps for sequencing of K182102-K082014-I763020 & R0063-F2621-I763020 & R0063-F2621-I0460 & R0063-F2621-K082014 & I0460-K176028
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We also made liquid cultures from the colonies that got PCR-confirmed.
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'''August 14'''
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Miniprepped the following parts: R0063-F2621-K082014-R0063-F2621-I763020 & R0063-F2621-I763020-R0063-F2621-K082014
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More digestions and ligations and transformations.
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'''August 15'''
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Colony pcr from yesterdays’ ligations.
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'''August 16'''
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Miniprepped the following parts: J23105-P0312 & K176028-I0460-K182102-K082014-R0062-I763020
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'''August 17'''
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Digestions, ligations, transformations.
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'''August 18'''
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Colony PCR
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'''August 23'''
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Performing a double ligation using R0040, J24675 & a linearized backbone.
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'''August 25'''
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Miniprepped R0040-J24675
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'''August 30'''
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Digestions, ligations, transformations.
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[https://2011.igem.org/Team:Wageningen_UR/Notebook/Proj1/July Previous Month]
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<!-- *** End of the "please do not edit" section. For the rest: Go ahead! :) *** -->
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[https://2011.igem.org/Team:Wageningen_UR/Notebook/Proj1/September Next Month]
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}}

Latest revision as of 18:00, 21 September 2011

Building a Synchronized Oscillatory System





August - Synchronized Oscillatory System

Aug.png

August 2

Colony PCR to verify ligations. Also some transformations.

August 3

Verifying parts using GFP and making liquid cultures from PCR- and GFP-confirmed colonies.

August 4

We isolated the following assembled parts: F2621-I763020 & R0062-I763020 & F2621-K082014-F2621-I763020 & F2621-I0460-F2621-I763020 & F2621-I0460-K318511 & R0062-I0460-K182102-K082014 & K182102-K082014-R0062-I763020

This day we discovered something horrible... Our sequencing shows that the F2621 part from the registry did not have the R0063 promoter in front of it so at this point we realized we had to redo a lot of our cloning...

August 5

We isolated the following assembled parts: K182102-C0061-B0015 & F2621-I0460

August 6

We isolated the assembled part: K182102-K082014 &

August 8

Digestions, ligations, transformations.

August 10

Digestions, ligations, transformations.

August 11

Yesterday’s gel showed that the digestions were a bit crappy so we redid all of them and then ligated and transformed again. Colony PCR of the transformants from the 10th of August.

August 12

Miniprepped the following parts: R0063-F2621-I0460 & R0063-F2621-I763020 & F2621-E0422-F2621-K082014 & R0063-F2621-K082014 & K176028-I0460 Digestions were started to then be ligated and used to transform E. coli.

August 13

PCR screen of the ligations from August 12th. Also minipreps for sequencing of K182102-K082014-I763020 & R0063-F2621-I763020 & R0063-F2621-I0460 & R0063-F2621-K082014 & I0460-K176028 We also made liquid cultures from the colonies that got PCR-confirmed.

August 14

Miniprepped the following parts: R0063-F2621-K082014-R0063-F2621-I763020 & R0063-F2621-I763020-R0063-F2621-K082014 More digestions and ligations and transformations.

August 15

Colony pcr from yesterdays’ ligations.

August 16

Miniprepped the following parts: J23105-P0312 & K176028-I0460-K182102-K082014-R0062-I763020

August 17

Digestions, ligations, transformations.

August 18

Colony PCR

August 23

Performing a double ligation using R0040, J24675 & a linearized backbone.

August 25

Miniprepped R0040-J24675

August 30

Digestions, ligations, transformations.


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