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Sigma WGA2

Library Preparation:

1. Add 2uL of 1x Library Preparation Buffer to each sample

2. Add 1uL of Library Stabilization Solution

3. Vortex thoroughly, consolidate by centrifugation and place in thermocycler at 95°C for 2 minutes

4. Cool the sample on ice for 5 minutes, consolidate by centrifugation and return the sample to ice

5. Add 1uL of Library Preparation Enzyme, vortex thoroughly, and centrifuge briefly

6. Place sample in a thermocycler and incubate as follows (listed as WGALIBRA)\

  • 16°C for 20 minutes
  • 24°C for 20 minutes
  • 37°C for 20 minutes
  • 75°C for 5 minutes
  • 4°C hold

7. Remove samples for thermocycler and centrifuge briefly. Amplify immediately

Amplification

8. A master mix may be prepared by adding the following reagents to the 15uL reaction in step 7. (when making the master mix only need to make enough for .25 extra. Ex. For 15 samples make enough mix for 15.25 samples)

  • 7.5uL of 10 Amplification Master
  • Mix 47.5uL of Nuclease Free Water
  • 5uL of WGA DNA Polymerase
  • Dispense 60uL per reaction

9. Vortex thoroughly, centrifuge briefly, and begin thermocycling at the following incubation times (listed as WGAAMP)

  • Initial Denaturation 95°C for 3 minutes
  • Perform 14 cycles as follows:
  1. Denature 94°C for 15 seconds
  2. Anneal/Extend 65°C for 5 minutes

After cycling is complete maintain the reaction of 4°C or store at -20 until ready for purification.

10. Proceed with Qiagen PCR Purification Kit