Shreffler:Notebook/retinoic acid trans signaling/2009/05/21

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(Autocreate 2009/05/21 Entry for Shreffler:Notebook/retinoic_acid_trans_signaling)
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==Entry title==
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*'''[[User:Neisha A. Rivers|Neisha A. Rivers]] 15:43, 3 June 2009 (EDT)''':
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* Insert content here...
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Prepare 1% Agarose Gel:
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(Note: Formula for 40 mls)
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1. Take out 0.4g of LE Agarose powder, and mix with 40 ml of TAE buffer
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2. Add 1 ul of ethidium bromide
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3. Heat in microwave for 30 sec to 1 min
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4. Allow to cool (so there are no bubbles), and place in the tray to dry
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5. Run between 80 to 100 Volts for approx 45 min to 1 hour
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----
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Calculations for Agarose Gel: (Want a concentration of 250 ng/ul)
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Loading Buffer = 5x concentration
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'''1x sample''': 189.7 ng/ul = 250/189.7 = 1.3 ul of DNA extract
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1.3 ul of DNA extract + 2 ul of loading buffer + 6.7 ul of H2O = 10 ul total
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'''2x sample''': 112.3 ng/ul = 250/112.3 = 2.2 ul of DNA extract
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2.2 ul of DNA extract + 2 ul of loading buffer + 5.8 ul of H2O = 10 ul total
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'''Plasmid sample''': 500 ng/ul = 250/500 = 0.5 ul of DNA extract
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0.5 ul of DNA extract + 2 ul of loading buffer + 7.5 ul of H2O = 10 ul total
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Revision as of 15:43, 3 June 2009

Retinoic Acid Trans Signaling Main project page
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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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