DspB Track
Marianne and Vicki
Gel Verification
- Verifying PCR products from last day (PCR-ing genomic preps)
- See Gel Verification Protocol in "iGEM Training Workshop Molecular Biology 2010"
Procedure:
- Prepare one 0.8% agarose gel
- Weigh out 0.8g of agarose and transfer to a 250mL flask
- Add 100mLs of 1x TBE buffer and swirl gently to disperse the agarose
- Microwave the mixture on high power until it boils and the agarose is completely dissolved.
- Allow the solution to cool to 60ºC by incubating in a 60ºC waterbath for about 10 minutes
- Add the appropriate (5uL per 50mL) amount of stain; in this case: add 10uL of GelStar
- Pour the cooled agarose into the plate with the appropriate number of combs
- Wait approximately 20 minutes
- Transfer 2uL of each PCR sample into new microfuge tubes
- Add 8uL of dH2O to each PCR sample
- Use quick and dirty way of loading gel:
- Pipet 2uL of DNA loading buffer for each of the samples onto parafilm
- Pipet 10uL of sample, then pipet the DNA loading buffer (2uL) for a total of 12uL, then load onto gel
Note: Used 100bp DNA ladder, a 1 in 20 dilution
Machine Conditions: 100V, 45 minutes, 1xTBE buffer
Gel arrangement:
Table 1. Arrangement of gel
| A | B | C | D | E | F | G | H | I | J | 100bp DNA ladder | K | L | M | N | O | P | Q | R | S |
Note: May have added more water in sample C -MP
Results:
Lab Stuff
- Autoclaved 10uL x2 boxes of pipette tips, microcentrifuge tubes 1.7mL x2 bins
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iGEM Project name 1
