We used cells isolated from tissue by laser microdissection to perform whole genome amplification (WGA) and then array comparative genome hybridization (aCGH). However, the results are no so good. Detection pVal for more than half of oligonucleotide probes does not reach significance.
Could you please provide me with information concerning WGA kits, which you used for preparation of DNA for aCGH? We used Repli-g Mini Kit (Qiagen).
I've tried this a couple of times in the past and it's always been sub-optimal. The problem seems to be non-uniform amplification of the gDNA making your array very wavy. If there's a particular locus you are interested in, then perhaps FISH or digital PCR might give you better answers? Some old colleagues of mine got some pretty nice copy number data from their laser microdissection/digital PCR setup.
http://www.ncbi.nlm.nih.gov/pubmed/19927313
http://www.ncbi.nlm.nih.gov/pubmed/20299530
Dear Scott, thanks for your suggestions! Now I am using PicoPLEX™ WGA Kit from Rubicon and I have a good results.
- Badges
- Science topic
- Similar topics
- Biological Science
- Microbiology
- Virology
- Virus