07/08/13

From 2013.igem.org

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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.
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.