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Revision as of 21:11, 2 August 2012 (view source) Vadeo (Talk | contribs) ← Older edit		Revision as of 21:11, 2 August 2012 (view source) Vadeo (Talk | contribs) Newer edit →
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	<h1>Week 7: 23rd July - 29th July</h1>		<h1>Week 7: 23rd July - 29th July</h1>
-	<h2>Monday, 23rd June</h2>	+	<h2>Monday, 23rd July</h2>
	<h3>Cloning:</h3>		<h3>Cloning:</h3>
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	<table/>		<table/>
-	<h2>Tuesday, 24th June</h2>	+	<h2>Tuesday, 24th July</h2>
	<h3>Plasmid Purification:</h3>		<h3>Plasmid Purification:</h3>
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	Gel 0,8%		Gel 0,8%
-	<h2>Thursday, 26th June</h2>	+	<h2>Thursday, 26th July</h2>
	Test of auxin toxicity in tadpodes.		Test of auxin toxicity in tadpodes.
-	<h2>Friday, 27th June</h2>	+	<h2>Friday, 27th July</h2>
	<ul>		<ul>

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Revision as of 21:11, 2 August 2012

Weeks

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

Week 7: 23rd July - 29th July

Monday, 23rd July

Cloning:

4 different clones pCS2+ RFP => incubation in LB medium overnight at 37 degree celsius.

PCR:

PCR of pCS2+:

Reactants	Volumes (µl)
GC Buffer	10
dNTPs	1
DNA	1
H2O	32,5
Primers (FW and RV)	2,5 each
fusion polymerase	0,5

Tuesday, 24th July

Plasmid Purification:

On the pCS2+ RFP clones incubated the day before:

DNA Concentration

Tube	Concentration (ng.uL-1)
1	144,2
2	161,9
3	184,9
4	120,7

Digestion:

Plasmid digestion

Tube	V DNA (uL)	V SpeI (uL)	V EcoRI (uL)	V buffer (uL)	V H2O (uL)
1	7	1	1	2	9
2	6,2	1	1	2	9,8
3	5,4	1	1	2	10,6
4	8,3	1	1	2	7,7

Gel migration:

Gel 0,8%

Thursday, 26th July

Test of auxin toxicity in tadpodes.

Friday, 27th July

• PCR of TirI: One PCR with primers TirI FW + Sdm RV / One PCR with primers TirI RV + Sdm FW
• Gel extraction of these PCR
• New PCR with the mix of the two PCR products + primers TirI FW and RV