Team:Paris/August 12

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Revision as of 09:30, 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	L128.1	pFlgA-RFP D132 (FI) - D136 (FV)
	L128.2	pFlgA-RFP D132 (FI) - D136 (FV)
	L128.3	pFlgA-RFP D132 (FI) - D136 (FV)
	L128.4	pFlgA-RFP D132 (FI) - D136 (FV)
MP145	L129.1	pFlgB-RFP D133 (FI) - D136 (FV)
	L129.2	pFlgB-RFP D133 (FI) - D136 (FV)
	L129.6	pFlgB-RFP D133 (FI) - D136 (FV)
	L129.7	pFlgB-RFP D133 (FI) - D136 (FV)
MP146	L130.1	pFlhB-RFP D134 (FI) - D136 (FV)
	L130.2	pFlhB-RFP D134 (FI) - D136 (FV)
	L130.7	pFlhB-RFP D134 (FI) - D136 (FV)
	L130.8	pFlhB-RFP D134 (FI) - D136 (FV)

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	127	1.67
MP145	166	1.66
MP146	163	1.57
MP147	154	1.63
MP148	136	1.67
MP149	151	1.63
MP150	163	1.67
MP151	188	1.57
MP152	298	1.70
MP153	142	1.64
MP154	170	1.52
MP155	143	1.55

New PCR screening with the right pimers

Transformants with pFlgA, pFlgB and pFlhB cloned into J61002 are analysed by PCR but this time with the right primers (O18 and 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 213: / Line 213: @@
 |align="center"|positive control
 |align="center"|negative control
-|colspan="4"|L128 (pFlgA)
+|colspan="4"|L128 ('''pFlgA''')
-|colspan="4"|L129 (pFlgB)
+|colspan="4"|L129 ('''pFlgB''')
-|colspan="4"|L130 (pFlhB)
+|colspan="4"|L130 ('''pFlhB''')
 |align="center"|100 bp DNA ladder
 |-
@@ Line 251: / Line 251: @@
 |-
 |align="center"|'''expected size'''
-|align="center"|ladder
+|align="center"|
 |align="center"|1161 bp
 |align="center"|0 bp
@@ Line 257: / Line 257: @@
 |colspan="4"|1339 bp
 |colspan="4"|1338 bp
-|align="center"|ladder
+|align="center"|
+|-
+|align="center"|'''measured size'''
+|align="center"|
+|align="center"|1,2 kb
+|align="center"|0 kb
+|align="center"|1,2 kb
+|align="center"|1,2 kb
+|align="center"|1,3 kb
+|align="center"|1,3 kb
+|align="center"|1,2 kb
+|align="center"|0 kb
+|align="center"|1,4 kb
+|align="center"|1,4 kb
+|align="center"|1,2 kb
+|align="center"|1,2 kb
+|align="center"|1,2 kb
+|align="center"|1,2 kb
+|align="center"|
 |}

Team:Paris/August 12

From 2008.igem.org

Revision as of 09:30, 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 pimers

Electrophoresis