Team:Paris/August 16

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Revision as of 14:53, 17 August 2008

Analysis of the transformation we did yesterday

L143, L144, T1 and T2 showed no colonies. The positive control with pUC19 worked well.
We suppose that the ligation did not work. We will do it again today.

for Kok-Phen Ligations

Ligation

Cleaning of the DNA after the digestion

We used the QIAcube to wash the DNA, following the standard protocol.

Measure of DNA concentration of the digestion products

We used the biophotometer.
Settings:

10 µL of template DNA in 50 µL of pure water
Blank : 10 µL of EB buffer in 50 µL of water.

Digestion name	What's in ?	DNA concentration (ng/µL)
D 149	pfliL	2
D 150	pfliL	3
D 151	pSB3K3	12
D 152	pSB3K3	20
D 153	gene flhDC	7
D 154	gene flhDC	12
D 155	pflhDC	16
D 136	j61002	12
D 145	pSB1A2	21

List of ligations

We ligated the DNA following the standard protocol.
T1, T2, T3 and T4 are the autoligation controls for L 143, L144, L145 and L147

Ligation name	Insert name	V_insert µL	Vector name	V_Vector µL
L 143 (Kan)	D 149 (pfliL)	8	D 137 (pSB3K3)	4
L 144 (Kan)	D 150 (pfliL)	5	D 152 (pSB3K3)	2.5
L 145 (Amp)	D 153 (g flhDC)	10	D 145 (pSB1A2)	2.5
L 146 (Amp)	D 154 (g flhDC)	5	D 145 (pSB1A2)	2.5
L 147 (Amp)	D 155 (p flhDC)	1	D 136 (J61002)	4

Transformation

Transformation protocol

Name	Description	Antibio
Ligations
L150	D105(BV) - D146(BI) pLas - strongest rbs-TetR-GFP tripart	Amp
L151	D147(FI) - D125 (FV) strongest rbs-LasR activator with LVA - Double terminator	Kan
Controls
C1	D105	Amp
C2	D125	Kan
Positive Control	pUC19	Amp

@@ Line 4: / Line 4: @@
 L143, L144, T1 and T2 showed no colonies. The positive control with pUC19 worked well.
 <br>We suppose that the ligation did not work. We will do it again today.
+==for Kok-Phen '''Ligations'''==
 =='''Ligation'''==