Download qBiomarker Somatic Mutation PCR Array System

Appendix A: Whole Genome Amplification of
Genomic DNA
Whole genome amplification (WGA) can dramatically reduce the required
amount of starting material. This protocol is for amplifying DNA from fresh or
frozen tissue samples for subsequent use with qBiomarker Somatic Mutation
PCR Arrays.
Note: The following procedure is a quick setup guide for whole genome
amplification using the REPLI-g UltraFast Mini Kit. For detailed instructions, refer
to the REPLI-G UltraFastMini Handbook.
Important points before starting

WGA is intended for fresh or frozen cell and tissue samples that contain
5–10 ng genomic DNA. If 200–500 ng genomic DNA is extracted from
fresh tissue and is of high quality, it is not necessary to perform WGA (see
“Important Notes”, page 16).

WGA is not recommended for DNA extracted from FFPE sections.
Procedure
A1.
Prepare sufficient Buffer D1 for the total number of WGA reactions,
including a wild-type control sample, according to Table 16.
Table 16. Preparation of Buffer D1
Component
Volume
Reconstituted Buffer DLB
5 µl
Nuclease-free water
35 µl
Total volume*
40 µl
* The total volume is sufficient for 40 samples.
A2.
Prepare sufficient Buffer N1 for the total number of WGA
reactions, including a wild-type control sample, according to Table
17.
qBiomarker Somatic Mutation PCR Handbook 08/2012
43