07/08/13
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+ | <div style="font-family:arial;padding:5px;border-radius:5px;border:5px solid #FF2800;"> | ||
+ | {| style="color:#87EA00;background-color:#FFFFFF;" cellpadding="2" cellspacing="2" border="0" bordercolor="#000000" width="100%" align="center" | ||
+ | !align="center"|[[Team:Leicester|Home]] | ||
+ | !align="center"|[[Team:Leicester/Team|Team]] | ||
+ | !align="center"|[https://igem.org/Team.cgi?year=2013&team_name=Leicester Official Team Profile] | ||
+ | !align="center"|[[Team:Leicester/Project|Project]] | ||
+ | !align="center"|[[Team:Leicester/Parts|Parts Submitted to the Registry]] | ||
+ | !align="center"|[[Team:Leicester/Modeling|Modeling]] | ||
+ | !align="center"|[[Team:Leicester/Notebook|Notebook]] | ||
+ | !align="center"|[[Team:Leicester/Safety|Safety]] | ||
+ | !align="center"|[[Team:Leicester/Attributions|Attributions]] | ||
+ | |} | ||
+ | </div> | ||
+ | |||
+ | |||
+ | ==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. | ||
+ | |||
{|border=1 | {|border=1 | ||
|Sample||Concentration (ng/ul)||260/280||260/230 | |Sample||Concentration (ng/ul)||260/280||260/230 | ||
Line 4: | Line 25: | ||
|1kb||534||1.82||1.95 | |1kb||534||1.82||1.95 | ||
|- | |- | ||
- | |1||52.7||1.77||2.05 | + | |1 5/45||52.7||1.77||2.05 |
|- | |- | ||
- | |2||4.6||1.46||1.88 | + | |2 5/45||4.6||1.46||1.88 |
|- | |- | ||
- | |3||0.1||0.24||-0.29 | + | |3 5/45||0.1||0.24||-0.29 |
|- | |- | ||
- | |Overnight | + | |Overnight* |
|- | |- | ||
|1in 10||24.8||1.8||2.42 | |1in 10||24.8||1.8||2.42 | ||
Line 20: | Line 41: | ||
|Fresh stock | |Fresh stock | ||
|- | |- | ||
- | | | + | |1s||5341.8||1.78||2.15 |
|- | |- | ||
- | | | + | |2s||440.0||1.80||2.23 |
|- | |- | ||
- | | | + | |3s||45.1||1.83||2.26 |
|} | |} | ||
+ | |||
+ | *concentration of DNA was measured using nanodrop. | ||
+ | |||
+ | *Dilutions left on the bench overnight. | ||
+ | |||
+ | == Gel electrophoresis of Dilluted Herring sperm DNA== | ||
+ | [[File: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. |
Latest revision as of 10:18, 8 August 2013
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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.
Sample | Concentration (ng/ul) | 260/280 | 260/230 |
1kb | 534 | 1.82 | 1.95 |
1 5/45 | 52.7 | 1.77 | 2.05 |
2 5/45 | 4.6 | 1.46 | 1.88 |
3 5/45 | 0.1 | 0.24 | -0.29 |
Overnight* | |||
1in 10 | 24.8 | 1.8 | 2.42 |
1 | 2.1 | 1.67 | 22.44 |
2 | 0.8 | 2.87 | -3.99 |
Fresh stock | |||
1s | 5341.8 | 1.78 | 2.15 |
2s | 440.0 | 1.80 | 2.23 |
3s | 45.1 | 1.83 | 2.26 |
- concentration of DNA was measured using nanodrop.
*Dilutions left on the bench overnight.
Gel electrophoresis of Dilluted Herring sperm DNA
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.