Line 392: | Line 392: | ||
</div></div></div> | </div></div></div> | ||
− | |||
− | |||
Line 402: | Line 400: | ||
<input type="button" style="height: 50px; width: 50%; BACKGROUND-COLOR: #3399FF; font-size:25; color:black;" onclick="showSpoiler(this);" value="DNA Purification" /> | <input type="button" style="height: 50px; width: 50%; BACKGROUND-COLOR: #3399FF; font-size:25; color:black;" onclick="showSpoiler(this);" value="DNA Purification" /> | ||
<div class="inner" style="display:none;"> | <div class="inner" style="display:none;"> | ||
− | |||
− | |||
<br><br> | <br><br> | ||
<p align="center"><b>1. Depending on the PCR product</b> <p> | <p align="center"><b>1. Depending on the PCR product</b> <p> | ||
Line 426: | Line 422: | ||
<b>2. Binding of DNA </b> | <b>2. Binding of DNA </b> | ||
− | <br> <br><div style="text- | + | <br><br><div style="text-align: justify; margin-left:40px"> |
− | 1. Insert SV Minicolumn into Collection Tube.< | + | 1. Insert SV Minicolumn into Collection Tube.<br> |
− | + | 2. Transfer dissolved gel mixture or prepared PCR product to the Minicolumn assembly. Incubate at room temperature for 1 minute. | |
− | 2. Transfer dissolved gel mixture or prepared PCR product to the Minicolumn assembly. | + | <br> |
− | at room temperature for 1 minute. | + | 3. Centrifuge at 16,000 ×g for 1 minute. Discard flowthrough and reinsert Minicolumn |
− | < | + | into Collection Tube.<br> |
− | 3. Centrifuge at 16,000 ×g for 1 minute. | + | |
− | into Collection Tube.< | + | |
− | + | ||
4. Heat NE-buffer to 70 °C.</div> | 4. Heat NE-buffer to 70 °C.</div> | ||
<br> | <br> | ||
<b>3. Washing </b> | <b>3. Washing </b> | ||
− | <br> <br><div style="text- | + | <br><br><div style="text-align: justify; margin-left:40px"> |
1. Add 700 μl Membrane Wash Solution (ethanol added). Centrifuge at 16,000 × g for 1 minute. | 1. Add 700 μl Membrane Wash Solution (ethanol added). Centrifuge at 16,000 × g for 1 minute. | ||
− | + | Discard flowthrough and reinsert Minicolumn into Collection Tube.<br> | |
− | + | ||
− | 2. Repeat Step 4 with 500 μl Membrane Wash Solution. Centrifuge at 16,000 × g for 5 minutes.< | + | 2. Repeat Step 4 with 500 μl Membrane Wash Solution. Centrifuge at 16,000 × g for 5 minutes.<br> |
− | + | ||
3. Empty the Collection Tube and recentrifuge the column assembly for 1 minute with the | 3. Empty the Collection Tube and recentrifuge the column assembly for 1 minute with the | ||
− | + | microcentrifuge lid open(or off to allow evaporation of any residual ethanol.</div> | |
<br> | <br> | ||
<b>4. Elution</b> | <b>4. Elution</b> | ||
− | <br> <br><div style="text- | + | <br><br><div style="text-align: justify; margin-left:40px"> |
− | 1. Carefully transfer Minicolumn to a clean 1.5 ml microcentrifuge tube.< | + | 1. Carefully transfer Minicolumn to a clean 1.5 ml microcentrifuge tube.<br> |
− | + | ||
− | 2. Add 50 μl of Nuclease-Free Water to the Minicolumn. Incubate at room temperature for 1 minute. | + | 2. Add 50 μl of Nuclease-Free Water to the Minicolumn. Incubate at room temperature for 1 minute. Centrifuge at 16,000 × g for 1 minute. <br> |
− | + | ||
− | 3. Discard Minicolumn and measure the concentration.< | + | 3. Discard Minicolumn and measure the concentration.<br> |
− | + | ||
4. Store DNA at 4°C or –20°C.</div> | 4. Store DNA at 4°C or –20°C.</div> | ||
<br> | <br> |
Revision as of 16:30, 31 October 2017
Our research work
We are describing our research work. Below you can find the protocols we used.
Protocols
Main sponsors: