22/08/13

From 2013.igem.org

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Revision as of 13:54, 22 August 2013

Measuring the concentrations of isolated P. putida DNA from the previous day

Measuring concentration using nanodrop
Blanking against buffer

Sample nr.	Concentration ng/ul	260/280	260/230
2	44	1.8	1.15
4	30.6	1.8	1.29
6	42.3	1.81	1.05
8	35.6	1.85	1.26
10	38.2	1.84	1.28
12	40.6	1.83	1.45
14	39.9	1.88	1.47
16	18.5	1.17	0.67

Preparing samples to run on 1% gel

500ng of each of the isolated P. putida DNA sample is run on gel
Volumes are adjusted to 20ul per well accordingly (includes DNA, sterile H2O and OG)
5ul of orange G dye is added to each sample
5ul of 1kb ladders are used
For sheared DNA, dilutions were loaded
- 1ul of DNA was diluted in 9ul of 1xTE buffer to make up 10x dilution
- 1ul from 10x dilution was put in 9ul of 1xTE buffer to make up 100x dilution
Dilutions were nanodropped and the concentration for 100 fold dilution was 121 ng/ul
200ng of herring sperm DNA was run on gel
One lane was left for control, non-sheared DNA, also 100 fold dilution

Running gel of P. putida DNA and Herring sperm DNA

Samples in lanes goes as follows: 1 Kb ladder, 2, 4, 6, 8, 10, 12, 14, 16, 1 kb ladder, 100x dilution of sheared herring sperm DNA, control 100x diluted herring sperm DNA

Revision as of 13:54, 22 August 2013 (view source) AliceHaworth (Talk \| contribs) (→Running sonicated herring sperm on gel) ← Older edit		Revision as of 13:54, 22 August 2013 (view source) AliceHaworth (Talk \| contribs) (→Running sonicated herring sperm on 0.8% gel) Newer edit →
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	Samples in lanes goes as follows: 1 Kb ladder, 2, 4, 6, 8, 10, 12, 14, 16, 1 kb ladder, 100x dilution of sheared herring sperm DNA, control 100x diluted herring sperm DNA		Samples in lanes goes as follows: 1 Kb ladder, 2, 4, 6, 8, 10, 12, 14, 16, 1 kb ladder, 100x dilution of sheared herring sperm DNA, control 100x diluted herring sperm DNA

-	==Running sonicated herring sperm on 0.8% gel==	+	==Running sonicated herring sperm DNA on 0.8% gel==