( ng column in a new 2 ml coll...)Protocol for GenomePlex Whole Genome Amplification from Formalin-Fixed Parrafin-Embedded (FFPE) tissue

ng column in a new 2 ml collection tube.
19. Elute DNA. Pipette 200 l of the Elution Solution directly into the center of the binding column and incubate at room temperature for 5 minutes. Centrifuge for 1 minute at >6,500g to elute the DNA.
20. Store DNA samples at -20C.

Note: This protocol can be performed without usage of xylene starting with step number 10. As a result of omitting xylene treatment step the amount of DNA will decrease by approximately 50% when compared to the protocol carried with a xylene step.

V. Protocol for GenomePlex Whole Genome Amplification from FFPE tissue
Performed with GenomePlex Whole Genome Amplification Kit (WGA1)
Fragmentation
1. Prepare DNA solution of 1 ng/l from the FFPE DNA
2. Add 1 l of 10X Fragmentation Buffer to 10 l DNA (1 ng/l) in a PCR tube.
3. Place the tube in a thermal cycler at 95 C for EXACTLY 4 minutes. Note, the incubation is time sensitive and any deviation may alter results.
4. Immediately cool the sample on ice and centrifuge briefly.

Library Preparation
5. Add 2 l of 1x Library Preparation Buffer.
6. Add 1 l of Library Stabilization Solution.
7. Mix thoroughly and place in thermal cycler at 95 C for 2 minutes.
8. Cool the sample on ice and centrifuge briefly.
9. Add 1 l Library Preparation Enzyme, mix thoroughly, and centrifuge briefly.
10. Place sample in thermal cycler and incubate as follows:
16 C for 20 minutes
24 C for 20 minutes
37 C for 20 minutes
75 C for 5 minutes
4 C hold
11. Remove samples from thermal cycler and centrifuge briefly. Samples
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