07/08/13

From 2013.igem.org

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(Dillution of Herring sperm DNA)
 
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{| style="color:#87EA00;background-color:#FFFFFF;" cellpadding="2" cellspacing="2" border="0" bordercolor="#000000" width="100%" align="center"
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!align="center"|[[Team:Leicester|Home]]
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!align="center"|[[Team:Leicester/Team|Team]]
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!align="center"|[https://igem.org/Team.cgi?year=2013&team_name=Leicester Official Team Profile]
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!align="center"|[[Team:Leicester/Project|Project]]
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!align="center"|[[Team:Leicester/Parts|Parts Submitted to the Registry]]
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!align="center"|[[Team:Leicester/Modeling|Modeling]]
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!align="center"|[[Team:Leicester/Notebook|Notebook]]
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!align="center"|[[Team:Leicester/Safety|Safety]]
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!align="center"|[[Team:Leicester/Attributions|Attributions]]
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|}
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==Dillution of Herring sperm DNA==
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*Dilutions were done in 3 sets:
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**Samples 1-3 were diluted in 90ul of water, where 10ul of stock DNA was added from an overnight dilution of 100um of Herring sperm DNA in 900ul of water.
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**Samples 1s-3s were also diluted in 90ul of water, except the first 10um of DNA was taken from an undiluted stock Herring sperm DNA.
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**Samples 1 5/45- 3 5/45 were from a known concentration of DNA stock, where 5ul of DNA was added to 45ul of water.
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{|border=1
{|border=1
|Sample||Concentration (ng/ul)||260/280||260/230
|Sample||Concentration (ng/ul)||260/280||260/230
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|1kb||534||1.82||1.95
|1kb||534||1.82||1.95
|-
|-
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|1||52.7||1.77||2.05
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|1 5/45||52.7||1.77||2.05
|-
|-
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|2||4.6||1.46||1.88
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|2 5/45||4.6||1.46||1.88
|-
|-
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|3||0.1||0.24||-0.29
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|3 5/45||0.1||0.24||-0.29
|-
|-
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|Overnight
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|Overnight*
|-
|-
|1in 10||24.8||1.8||2.42
|1in 10||24.8||1.8||2.42
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|Fresh stock
|Fresh stock
|-
|-
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|1||5341.8||1.78||2.15
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|1s||5341.8||1.78||2.15
|-
|-
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|2||440.0||1.80||2.23
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|2s||440.0||1.80||2.23
|-
|-
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|3||45.1||1.83||2.26
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|3s||45.1||1.83||2.26
|}
|}
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*concentration of DNA was measured using nanodrop.
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*Dilutions left on the bench overnight.
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== Gel electrophoresis of Dilluted Herring sperm DNA==
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[[File:iGEM_DNA_070813.jpg]]
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Lanes are: marker, 1 5/45, 2 5/45, 3 5/45, 1, 2, 3, 1s, 2s, 3s, lambda NT, lambda
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Results are as expected. Lanes show that samples 1, 2 and 3 were degraded overnight, suggesting nuclease activity. Samples 1s, 2s and 3s are the fresh Herring sperm stock and show no degradation.

Latest revision as of 10:18, 8 August 2013

Home Team Official Team Profile Project Parts Submitted to the Registry Modeling Notebook Safety Attributions


Dillution of Herring sperm DNA

  • Dilutions were done in 3 sets:
    • Samples 1-3 were diluted in 90ul of water, where 10ul of stock DNA was added from an overnight dilution of 100um of Herring sperm DNA in 900ul of water.
    • Samples 1s-3s were also diluted in 90ul of water, except the first 10um of DNA was taken from an undiluted stock Herring sperm DNA.
    • Samples 1 5/45- 3 5/45 were from a known concentration of DNA stock, where 5ul of DNA was added to 45ul of water.
SampleConcentration (ng/ul)260/280260/230
1kb5341.821.95
1 5/4552.71.772.05
2 5/454.61.461.88
3 5/450.10.24-0.29
Overnight*
1in 1024.81.82.42
12.11.6722.44
20.82.87-3.99
Fresh stock
1s5341.81.782.15
2s440.01.802.23
3s45.11.832.26
  • concentration of DNA was measured using nanodrop.
*Dilutions left on the bench overnight.

Gel electrophoresis of Dilluted Herring sperm DNA

IGEM DNA 070813.jpg

Lanes are: marker, 1 5/45, 2 5/45, 3 5/45, 1, 2, 3, 1s, 2s, 3s, lambda NT, lambda

Results are as expected. Lanes show that samples 1, 2 and 3 were degraded overnight, suggesting nuclease activity. Samples 1s, 2s and 3s are the fresh Herring sperm stock and show no degradation.