Team:Paris/August 11

From 2008.igem.org

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==PCR==
==PCR==
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''We performed PCR on to amplify the sequence in order to have enough amount of DNA to carry out the following of our experiments.''
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=== '''PCR amplification''' ===
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==== '''Protocol''' ====
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* '''List of Oligos''' :
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{|
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|- style="background: #649CD7;"
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|- style="background: #649CD7; text-align: center;"
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|width=5%| Number
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|width=15%| Name
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|width=40%| Sequence
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|width=5%| Length
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|width=30%| Comments
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|- style="background: #dddddd;"
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| style="background: #D4E2EF;" | O110
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| FlhDC-Uri-F
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| GTTTCTTCGAATTCGCGGCCGCTTCTAGAGTTGTATGTGCGTGTAGTGACGAGTACAG
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| 58
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|
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|- style="background: #dddddd;"
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| style="background: #D4E2EF;" | O111
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| FlhDC-Total-F
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| GTTTCTTCGAATTCGCGGCCGCTTCTAGAGTCATTTTTGCTTGCTAGCGTACGGAAAA
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| 58
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|
==Culture of ligation transformants==
==Culture of ligation transformants==

Revision as of 09:37, 12 August 2008

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Contents

Transformation

Digestion

PCR

We performed PCR on to amplify the sequence in order to have enough amount of DNA to carry out the following of our experiments.


PCR amplification

Protocol

  • List of Oligos :
Number Name Sequence Length Comments
O110 FlhDC-Uri-F GTTTCTTCGAATTCGCGGCCGCTTCTAGAGTTGTATGTGCGTGTAGTGACGAGTACAG 58
O111 FlhDC-Total-F GTTTCTTCGAATTCGCGGCCGCTTCTAGAGTCATTTTTGCTTGCTAGCGTACGGAAAA 58

Culture of ligation transformants

  • 4 clones of each transformation were cultured in 7,5 mL LB + ampicilline. The colonies picked up were the rest of those already picked up from the transformation plate (for the PCR screening of August 8).
  • 37°C overnight
Ligation L128 L129 L130
Name pFlgA pFlgB pFlhB
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
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