Dear Li,

Its absolutely depends on your objective...why do you want only HPLC-MSP method?

As far as I know for candidate gene specific methylation quantitative techniques they includes:

1) Methyl light MSP (Sensitive one),

2) COBRA, Pyrosequencing and MASS Array MS technique (quantitative but not allele specific)

3) Bisulphite cloning+sequencing (Allele specific methylation)

Based on my personal experience I am very satisfied with Pyrosequncing and MASS Array....but they are bit expensive (but not if you are analyzing many samples > 50, as in my case)

Methyl light MSP is comparatively cheap one and works fine as well.

Hope this will be helpful...goodluck for your method selection...

Need any help for primer designing just contact me (in case you would like to try my recommended methods)

Dear Li,

SHOX2 is already established as a biomarker for lung cancer. There is a PCR assay available commercially available. Furthermore, 2 established assays can be found here:

http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0084225

http://www.ncbi.nlm.nih.gov/pubmed/22555092

I hope this helps.

Best,

Dimo

thank for sincere answer above.thanl Dimo Dietrich  and Tushar Tomar