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Revision as of 13:14, 21 July 2015
Protocols
Agarose Gel
Materials
• 1X TAE
• Agarose
• Gel Red
• DNA samples
• 6X loading dye
• Nuclease free water
Procedure
Prepare 1.2% agarose gel for small fragments and 3% agarose gel for large fragments
• Mix 50 mL 1X TAE and 0.6 g (1.2%) or 1.5 g (3%) agarose
• Melt in microwave until agarose has melted (about 50 seconds)
• Add 1.3 μL (1.2%) or 1.5 μL (3%) Gel Red
• Pour solution into agarose gel mold with comb
• Let set for 20 minutes or until solid
• Place gel in 1X TAE and remove comb
• Load samples of 200 ng (or 2 μL) DNA mixed with 2 μL 6X loading dye and nuclease free water up to 12 μL
• Run gel at 100-120 Volts for 40-50 minutes (1.2%) or 80 Volts for 2 hours (3%)
• Take a picture of the gel at the UV detector
Amino acid solution
Materials
• Histidine-Hcl
• Uracil
• Leucine
• Tryptophan
Procedure
| Stock concentration | Final concentration | Total quantity for 50 mL |
|---|---|---|
| 100 mM Histidine-Hcl (209 g/mol) | 20.9 g/L | 0.418 g |
| 20 mM Uracil (112 g/mol) | 2.24 g/L | 0.0448 g |
| 100 mM Leucine (131 g/mol) | 13.1 g/L | 0.262 g |
| 40 mM Tryptophan (204 g/mol) | 8.16 g/L | 0.1632 g |
• Filter and sterilize solutions
•Add 8 mL per liter of selective medium or spread 500 μL on a selective plate
Section 3
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Section 4-1
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Section 4-2
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