Shreffler:Notebook/retinoic acid trans signaling/2009/05/21: Difference between revisions
(Autocreate 2009/05/21 Entry for Shreffler:Notebook/retinoic_acid_trans_signaling) |
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== | *'''[[User:Neisha A. Rivers|Neisha A. Rivers]] 15:43, 3 June 2009 (EDT)''': | ||
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 | |||
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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 | |||
Revision as of 12:43, 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
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