Team:Queens Canada/Notebook/Protocols/Rehydration

From 2011.igem.org

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<regulartext> 2. Using sterile technique, add the correct volume of ddH2O to the dry primers such that they make a 100μM solution (consult the shipping invoice to see what the volume of water should be). If the volume specified by the invoice is over 900μL, add half the specified volume (to make a 200μM solution). Label the top of the tube with the concentration of the primer.    </regulartext> <br>
<regulartext> 2. Using sterile technique, add the correct volume of ddH2O to the dry primers such that they make a 100μM solution (consult the shipping invoice to see what the volume of water should be). If the volume specified by the invoice is over 900μL, add half the specified volume (to make a 200μM solution). Label the top of the tube with the concentration of the primer.    </regulartext> <br>
<regulartext> 3. Briefly vortex the tube to ensure that all the DNA has dissolved (make sure there are no white flecks remaining in the bottom of the tube).  </regulartext> <br>
<regulartext> 3. Briefly vortex the tube to ensure that all the DNA has dissolved (make sure there are no white flecks remaining in the bottom of the tube).  </regulartext> <br>
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<regulartext> 4. Let the tube sit for 1-3 minutes to ensure that the primers have completely dissolved. It is convenient to label your tubs during this time.  
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<regulartext> 4. Let the tube sit for 1-3 minutes to ensure that the primers have completely dissolved. It is convenient to label your tubs during this time. <br>
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5. Briefly vortex again.   
+
5. Briefly vortex again.  <br>
-
6. Aliquot out the correct volume of the concentrated (100 or 200μM) primer solution into an eppendorf tube and dilute it to 100μL solution at the concentration of 10μM (if your concentrated primer solution is 100μM, use 10μL or the concentrated primer and 90μL of water to make your 10μM primer solution).  
+
6. Aliquot out the correct volume of the concentrated (100 or 200μM) primer solution into an eppendorf tube and dilute it to 100μL solution at the concentration of 10μM (if your concentrated primer solution is 100μM, use 10μL or the concentrated primer and 90μL of water to make your 10μM primer solution).  
  </regulartext> <br>
  </regulartext> <br>

Revision as of 12:42, 19 September 2011

Queen's
rehydration
Rehydration- iGEM Standard Parts

Storage and Labelling
- Rehydrate primers just before they are needed for PCR
- Store primers (both concentrations) in the -20C freezer

Materials
- Dry primers
- ddH2O

Procedure
1. Ensure that you know what the primer that you are rehydrating is for so that you can label it properly.
2. Using sterile technique, add the correct volume of ddH2O to the dry primers such that they make a 100μM solution (consult the shipping invoice to see what the volume of water should be). If the volume specified by the invoice is over 900μL, add half the specified volume (to make a 200μM solution). Label the top of the tube with the concentration of the primer.
3. Briefly vortex the tube to ensure that all the DNA has dissolved (make sure there are no white flecks remaining in the bottom of the tube).
4. Let the tube sit for 1-3 minutes to ensure that the primers have completely dissolved. It is convenient to label your tubs during this time.
5. Briefly vortex again.
6. Aliquot out the correct volume of the concentrated (100 or 200μM) primer solution into an eppendorf tube and dilute it to 100μL solution at the concentration of 10μM (if your concentrated primer solution is 100μM, use 10μL or the concentrated primer and 90μL of water to make your 10μM primer solution).

Rehydration- Primers

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