Team:Calgary/Notebook/Protocols/Process11
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<p>This protocol is utilized in accordance to the manufacturer's protocol from Omega E.Z.N.A (EaZy Nucleic Acid Isolation)</p> | <p>This protocol is utilized in accordance to the manufacturer's protocol from Omega E.Z.N.A (EaZy Nucleic Acid Isolation)</p> | ||
<ol> | <ol> | ||
<li> Place gel on the UV box</li> | <li> Place gel on the UV box</li> | ||
- | <li> Carefully extract the fragment suspended in the gel | + | <li> Carefully extract the fragment suspended in the gel </li> |
<li> Mass gel fragments </li> | <li> Mass gel fragments </li> | ||
- | <li> Place fragment into a 1.5 mL tube and add 4 µL of | + | <li> Place fragment into a 1.5 mL tube and add 4 µL of H<sub>2</sub>O </li> |
<li> Volume of water added to volume of gel is 200% however if fragment it small 1 mL of water will suffice </li> | <li> Volume of water added to volume of gel is 200% however if fragment it small 1 mL of water will suffice </li> | ||
- | <li> Remove | + | <li> Remove H<sub>2</sub>O</li> |
<li> Add equal amounts of H2O and Binding Buffer (XP2) to the gel</li> | <li> Add equal amounts of H2O and Binding Buffer (XP2) to the gel</li> | ||
<li> Incubate mixture at 55 degrees for 7 mins </li> | <li> Incubate mixture at 55 degrees for 7 mins </li> | ||
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<li> Discard the liquid </li> | <li> Discard the liquid </li> | ||
<li> Spin down the column at 13,000xg for 1 min to dry the column </li> | <li> Spin down the column at 13,000xg for 1 min to dry the column </li> | ||
- | <li> Elute in 50 µL of | + | <li> Elute in 50 µL of H<sub>2</sub>O and wait 1 min </li> |
<li> Spin down the column at 13,000xg for 1 min to dry the column </li> | <li> Spin down the column at 13,000xg for 1 min to dry the column </li> | ||
<li> Use a spectrophotometer to measure the concentration and the purity of your plasmid</li> | <li> Use a spectrophotometer to measure the concentration and the purity of your plasmid</li> | ||
</ol> | </ol> | ||
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+ | </html>}} |
Latest revision as of 04:24, 29 September 2011
Gel Extraction
This protocol is utilized in accordance to the manufacturer's protocol from Omega E.Z.N.A (EaZy Nucleic Acid Isolation)
- Place gel on the UV box
- Carefully extract the fragment suspended in the gel
- Mass gel fragments
- Place fragment into a 1.5 mL tube and add 4 µL of H2O
- Volume of water added to volume of gel is 200% however if fragment it small 1 mL of water will suffice
- Remove H2O
- Add equal amounts of H2O and Binding Buffer (XP2) to the gel
- Incubate mixture at 55 degrees for 7 mins
- Mix with vortex for 2 mins
- Place in the HiBind DNA Mini Column in the 2 mL tube
- Add 700 µL at 10,000xg for 1 min
- Discard liquid
- Add 300 µL Binding Buffer (XP2) into the HiBind DNA Mini Column and spin down at 10,000xg for 1 min
- Discard liquid
- Wash the column with 700 µL of SPW buffer with added ethanol and spin down at 10,000xg for 1 min
- Discard liquid
- Wash the column with 700 µL of SPW buffer again and spin down at 10,000xg for 1 min
- Discard the liquid
- Spin down the column at 13,000xg for 1 min to dry the column
- Elute in 50 µL of H2O and wait 1 min
- Spin down the column at 13,000xg for 1 min to dry the column
- Use a spectrophotometer to measure the concentration and the purity of your plasmid