Shreffler:Notebook/retinoic acid trans signaling/2009/05/21: Difference between revisions
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5. Run between 80 to 100 Volts for approx 45 min to 1 hour | 5. Run between 80 to 100 Volts for approx 45 min to 1 hour | ||
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0.5 ul of DNA extract + 2 ul of loading buffer + 7.5 ul of H2O = 10 ul total | 0.5 ul of DNA extract + 2 ul of loading buffer + 7.5 ul of H2O = 10 ul total | ||
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Agarose Gel Layout: | |||
1st well: 1 Kb Plus Ladder 250 ug (1.0 ul/ul) = 10 ul | |||
3rd well: 10 ul of 1x sample | |||
5th well: 10 ul of 2x sample | |||
7th well: Plasmid sample | |||
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Revision as of 12:46, 3 June 2009
 Retinoic Acid Trans Signaling
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Prepare 1% Agarose Gel: (Note: Formula for 40 mls) 1. Take out 0.4g of LE Agarose powder, and mix with 40 ml of TAE buffer 2. Add 1 ul of ethidium bromide 3. Heat in microwave for 30 sec to 1 min 4. Allow to cool (so there are no bubbles), and place in the tray to dry 5. Run between 80 to 100 Volts for approx 45 min to 1 hour Calculations for Agarose Gel: (Want a concentration of 250 ng/ul) Loading Buffer = 5x concentration 1x sample: 189.7 ng/ul = 250/189.7 = 1.3 ul of DNA extract 1.3 ul of DNA extract + 2 ul of loading buffer + 6.7 ul of H2O = 10 ul total 2x sample: 112.3 ng/ul = 250/112.3 = 2.2 ul of DNA extract 2.2 ul of DNA extract + 2 ul of loading buffer + 5.8 ul of H2O = 10 ul total Plasmid sample: 500 ng/ul = 250/500 = 0.5 ul of DNA extract 0.5 ul of DNA extract + 2 ul of loading buffer + 7.5 ul of H2O = 10 ul total Agarose Gel Layout: 1st well: 1 Kb Plus Ladder 250 ug (1.0 ul/ul) = 10 ul 3rd well: 10 ul of 1x sample 5th well: 10 ul of 2x sample 7th well: Plasmid sample
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