Team:Paris/August 12

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Revision as of 13:22, 13 August 2008

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Minipreps: Plasmid extraction

Protocol (see # 3) Experiments done by QIAcube

List of Minipreps

Name	Ligation	Biobricks	Description
MP144.1	L128.1	pFlgA-RFP D132 (FI) - D136 (FV)
MP144.2	L128.2
MP144.3	L128.3
MP144.4	L128.4
MP145.1	L129.1	pFlgB-RFP D133 (FI) - D136 (FV)
MP145.2	L129.2
MP145.6	L129.6
MP145.7	L129.7
MP146.1	L130.1	pFlhB-RFP D134 (FI) - D136 (FV)
MP146.2	L130.2
MP146.7	L130.7
MP146.8	L130.8

Glycerol Stocks

Protocol (see #2)

List of Stocks

Strain	Ligation	Biobricks	Description
S143.1	L128.1	pFlgA-RFP D132 (FI) - D136 (FV)
S143.2	L128.2
S143.3	L128.3
S143.4	L128.4
S144.1	L129.1	pFlgB-RFP D133 (FI) - D136 (FV)
S144.2	L129.2
S144.6	L129.6
S144.7	L129.7
S145.1	L130.1	pFlhB-RFP D134 (FI) - D136 (FV)
S145.2	L130.2
S145.7	L130.7
S145.8	L130.8

Results of the transformations we did yesterday

Concentration of the MiniPreps

Miniprep	Concentration (µg/mL)	Ratio 260/280
MP144.1	127	1.67
MP144.2	166	1.66
MP144.3	163	1.57
MP144.4	154	1.63
MP145.1	136	1.67
MP145.2	151	1.63
MP145.6	163	1.67
MP145.7	188	1.57
MP146.1	298	1.70
MP146.2	142	1.64
MP146.7	170	1.52
MP146.8	143	1.55

New PCR screening with the right primers

Transformants with pFlgA, pFlgB and pFlhB cloned into J61002 are analysed by PCR but this time with the right primers: VF2 (O18) and VR (O19).

template: bacteria from glycerol stock
positive control: J61002 (with ptet-mRFP)
negative control: no template

Electrophoresis

1% agarose gel
10 µL of each sample loaded

well n°

1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

sample

1 kb DNA ladder

positive control

negative control

L128 (pFlgA)

L129 (pFlgB)

L130 (pFlhB)

100 bp DNA ladder

clone n°

1

2

3

4

1

2

6

7

1

2

7

8

red fluorescence

yes

no

yes

no

yes

no

expected size

1161 bp

0 bp

1339 bp

1338 bp

measured size

1,2 kb

0 kb

1,2 kb

1,3 kb

1,2 kb

0 kb

1,4 kb

1,2 kb

@@ Line 121: / Line 121: @@
 |align="center"|'''Ratio 260/280'''
 |-
-|align="center"|MP144
+|align="center"|MP144.1
 |align="center"|127
 |align="center"|1.67
 |-
-|align="center"|MP145
+|align="center"|MP144.2
 |align="center"|166
 |align="center"|1.66
 |-
-|align="center"|MP146
+|align="center"|MP144.3
 |align="center"|163
 |align="center"|1.57
 |-
-|align="center"|MP147
+|align="center"|MP144.4
 |align="center"|154
 |align="center"|1.63
 |-
-|align="center"|MP148
+|align="center"|MP145.1
 |align="center"|136
 |align="center"|1.67
 |-
-|align="center"|MP149
+|align="center"|MP145.2
 |align="center"|151
 |align="center"|1.63
 |-
-|align="center"|MP150
+|align="center"|MP145.6
 |align="center"|163
 |align="center"|1.67
 |-
-|align="center"|MP151
+|align="center"|MP145.7
 |align="center"|188
 |align="center"|1.57
 |-
-|align="center"|MP152
+|align="center"|MP146.1
 |align="center"|298
 |align="center"|1.70
 |-
-|align="center"|MP153
+|align="center"|MP146.2
 |align="center"|142
 |align="center"|1.64
 |-
-|align="center"|MP154
+|align="center"|MP146.7
 |align="center"|170
 |align="center"|1.52
 |-
-|align="center"|MP155
+|align="center"|MP146.8
 |align="center"|143
 |align="center"|1.55

Team:Paris/August 12

From 2008.igem.org

Revision as of 13:22, 13 August 2008

Contents

Minipreps: Plasmid extraction

Glycerol Stocks

Results of the transformations we did yesterday

Concentration of the MiniPreps

New PCR screening with the right primers

Electrophoresis