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Team:Macquarie Australia/Protocols/RD
From 2012.igem.org
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<center><h1>Restriction Digest</h1></center> | <center><h1>Restriction Digest</h1></center> | ||
| - | <p>We prepared Master Mixes containing | + | <p>We prepared Master Mixes containing the following volumes,</p> |
<center><table><tr><td><table border="3" cellpadding="4" cellspacing="0"> | <center><table><tr><td><table border="3" cellpadding="4" cellspacing="0"> | ||
<tr><td colspan="2">EcoR1 + Spe1 Master Mix</td></tr> | <tr><td colspan="2">EcoR1 + Spe1 Master Mix</td></tr> | ||
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</table></td> | </table></td> | ||
<td><table border="3" cellpadding="4" cellspacing="0"> | <td><table border="3" cellpadding="4" cellspacing="0"> | ||
| - | <tr><td colspan="2"> | + | <tr><td colspan="2">Xba1 + Pst Master Mix</td></tr> |
<tr><td>Substrate</td><td>Volume (µL)</td></tr> | <tr><td>Substrate</td><td>Volume (µL)</td></tr> | ||
<tr><td>10x Buffer</td><td>20</td></tr> | <tr><td>10x Buffer</td><td>20</td></tr> | ||
| - | <tr><td> | + | <tr><td>Xba1</td><td>2.5</td></tr> |
| - | <tr><td> | + | <tr><td>Pst</td><td>2.5</td></tr> |
<tr><td>Water</td><td>75</td></tr> | <tr><td>Water</td><td>75</td></tr> | ||
<tr><td>Total</td><td>100</td></tr> | <tr><td>Total</td><td>100</td></tr> | ||
</table></td> | </table></td> | ||
<td><table border="3" cellpadding="4" cellspacing="0"> | <td><table border="3" cellpadding="4" cellspacing="0"> | ||
| - | <tr><td colspan="2">EcoR1 + | + | <tr><td colspan="2">EcoR1 + Pst Master Mix</td></tr> |
<tr><td>Substrate</td><td>Volume (µL)</td></tr> | <tr><td>Substrate</td><td>Volume (µL)</td></tr> | ||
<tr><td>10x Buffer</td><td>20</td></tr> | <tr><td>10x Buffer</td><td>20</td></tr> | ||
<tr><td>EcoR1</td><td>2.5</td></tr> | <tr><td>EcoR1</td><td>2.5</td></tr> | ||
| - | <tr><td> | + | <tr><td>Pst</td><td>2.5</td></tr> |
<tr><td>Water</td><td>75</td></tr> | <tr><td>Water</td><td>75</td></tr> | ||
<tr><td>Total</td><td>100</td></tr> | <tr><td>Total</td><td>100</td></tr> | ||
</table></td> | </table></td> | ||
| - | < | + | </table></center> |
| - | < | + | <h2>Protocol</h2> |
| - | < | + | <blockquote><ol><li>Mix 10 µL of the desired DNA/plasmid and 10 µL of the appropriate Master Mix in a PCR tube.</li> |
| - | < | + | <li>Using a thermocycler, incubate the mixture at 37°C for 30 minutes and then deactivate the enzymes by heating to 80°C for 20 minutes.</li> |
| - | + | <li>Store in a -20°C freezer until DNA required</li></blockquote></ol> | |
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</html> | </html> | ||
Latest revision as of 01:24, 26 September 2012
Restriction Digest
We prepared Master Mixes containing the following volumes,
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Protocol
- Mix 10 µL of the desired DNA/plasmid and 10 µL of the appropriate Master Mix in a PCR tube.
- Using a thermocycler, incubate the mixture at 37°C for 30 minutes and then deactivate the enzymes by heating to 80°C for 20 minutes.
- Store in a -20°C freezer until DNA required
