Team:Paris/August 12

From 2008.igem.org

(Difference between revisions)
(→New PCR screening with the right pimers)
(→New PCR screening with the right pimers)
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Transformants with pFlgA, pFlgB and pFlhB cloned into J61002 are analysed by PCR but this time with the right primers (O18 and O19).  
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
+
*template: bacteria from glycerol stock
*positive control: J61002 (with ptet-mRFP)
*positive control: J61002 (with ptet-mRFP)
*negative control: no template
*negative control: no template

Revision as of 12:26, 13 August 2008

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Contents

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) 		Part icon regulatory.pngPart icon reporter.png
L128.2 pFlgA-RFP
D132 (FI) - D136 (FV) 		Part icon regulatory.pngPart icon reporter.png
L128.3 pFlgA-RFP
D132 (FI) - D136 (FV) 		Part icon regulatory.pngPart icon reporter.png
L128.4 pFlgA-RFP
D132 (FI) - D136 (FV) 		Part icon regulatory.pngPart icon reporter.png
MP145 L129.1 pFlgB-RFP
D133 (FI) - D136 (FV) 		Part icon regulatory.pngPart icon reporter.png
L129.2 pFlgB-RFP
D133 (FI) - D136 (FV) 		Part icon regulatory.pngPart icon reporter.png
L129.6 pFlgB-RFP
D133 (FI) - D136 (FV) 		Part icon regulatory.pngPart icon reporter.png
L129.7 pFlgB-RFP
D133 (FI) - D136 (FV) 		Part icon regulatory.pngPart icon reporter.png
MP146 L130.1 pFlhB-RFP
D134 (FI) - D136 (FV) 		Part icon regulatory.pngPart icon reporter.png
L130.2 pFlhB-RFP
D134 (FI) - D136 (FV) 		Part icon regulatory.pngPart icon reporter.png
L130.7 pFlhB-RFP
D134 (FI) - D136 (FV) 		Part icon regulatory.pngPart icon reporter.png
L130.8 pFlhB-RFP
D134 (FI) - D136 (FV) 		Part icon regulatory.pngPart icon reporter.png

Glycerol Stocks

  • List of Stocks
Strain Ligation Biobricks Description
S143.1 L128.1 pFlgA-RFP
D132 (FI) - D136 (FV) 		Part icon regulatory.pngPart icon reporter.png
S143.2 L128.2
S143.3 L128.3
S143.4 L128.4
S144.1 L129.1 pFlgB-RFP
D133 (FI) - D136 (FV) 		Part icon regulatory.pngPart icon reporter.png
S144.2 L129.2
S144.6 L129.6
S144.7 L129.7
S145.1 L130.1 pFlhB-RFP
D134 (FI) - D136 (FV) 		Part icon regulatory.pngPart icon reporter.png
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 yes no no yes yes no no yes no no no
expected size 1161 bp 0 bp 1339 bp 1339 bp 1338 bp
measured size 1,2 kb 0 kb 1,2 kb 1,2 kb 1,3 kb 1,3 kb 1,2 kb 0 kb 1,4 kb 1,4 kb 1,2 kb 1,2 kb 1,2 kb 1,2 kb
Screening-PCR-080812.JPG
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