IGEM:Groningen/Notebook/iGEM 2011/2011/06/20

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(Autocreate 2011/06/20 Entry for IGEM:Groningen/Notebook/iGEM_2011)
Current revision (05:59, 21 September 2011) (view source)
(Entry title)
 
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==Entry title==
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==20-6-11==
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* Insert content here...
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<br>
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<br> PCRs with different samples, scheme:
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<br> Samples PhybB:          DNA TG1    3×TG1  Colony DH5alpha  3× colonies DH5alpha
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<br> pfu 10× buffer:          5μl        15μl        5μl                15μl
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<br> dNTPs 10mM:              1μl        3μl        1μl                  3μl
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<br> Forward Biobrick primer: 1μl        3μl        1μl                  3μl
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<br> Reverse Biobrick primer: 1μl        3μl        1μl                  3μl 
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<br> Pfu DNA polymerase:      1μl        3μl        1μl                  3μl
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<br> MQ water:                41μl        123μl    41μl                123μl
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<br>
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<br> PBAD and PlasI PCRs:
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<br> pfu 10× buffer:          5μl       
 +
<br> dNTPs 10mM:              1μl       
 +
<br> Forward Biobrick primer: 1μl         
 +
<br> Reverse Biobrick primer: 1μl           
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<br> Pfu DNA polymerase:      1μl       
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<br> MQ water:                41μl
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<br>
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<br>PCR conditions:
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<br>Preheated lid: 111°C
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<br>Denaturation: 95°C for 3 min.  
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<br>Cycle 33×:
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<br>denaturation: 95°C for 30s.  
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<br>annealing: 60°C for 30s.  
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<br>Extension: 72°C for 2.5 min.
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<br>Final extension: 72°C for 10 min.
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<br>Store infinite at 4°C
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<br>
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<br> Analyse samples on a 1% agarose gel
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<br> Clean up the DNA samples
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<br> work on human practices
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20-6-11



PCRs with different samples, scheme:
Samples PhybB: DNA TG1 3×TG1 Colony DH5alpha 3× colonies DH5alpha
pfu 10× buffer: 5μl 15μl 5μl 15μl
dNTPs 10mM: 1μl 3μl 1μl 3μl
Forward Biobrick primer: 1μl 3μl 1μl 3μl
Reverse Biobrick primer: 1μl 3μl 1μl 3μl
Pfu DNA polymerase: 1μl 3μl 1μl 3μl
MQ water: 41μl 123μl 41μl 123μl

PBAD and PlasI PCRs:
pfu 10× buffer: 5μl
dNTPs 10mM: 1μl
Forward Biobrick primer: 1μl
Reverse Biobrick primer: 1μl
Pfu DNA polymerase: 1μl
MQ water: 41μl

PCR conditions:
Preheated lid: 111°C
Denaturation: 95°C for 3 min.
Cycle 33×:
denaturation: 95°C for 30s.
annealing: 60°C for 30s.
Extension: 72°C for 2.5 min.
Final extension: 72°C for 10 min.
Store infinite at 4°C

Analyse samples on a 1% agarose gel
Clean up the DNA samples
work on human practices



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