bisulfite treatment for methylation-sensitive PCR

biomol at biomol at
Tue Nov 23 14:45:11 EST 2004

We are starting to work with methylation-sensitive PCR, for specific genes in leukemia. The protocol includes 
1- Bisulfite treatment (0,6%) plus Hydroquinona (0.4%) / NaOH (0.4%) on 1-3ug DNA (16-20 hs, 55ºC) 
2-Cleaning-up with Wizard DNA standard column 
3-ETOH+ NaAc precipitation 
4- PCR 

We would like to know if somebody has tried scaling-up bisulfite treatment, and which are the limiting factors, as we need to screen several genes and volumes are insufficient 
We also would appreciate indications about some cellular control genes that work well with this method. 

Thanks in advance! 

Esteban Braggio 
Laboratório de Biologia Molecular 
National Institute of Cancer 
Rio de Janeiro, Brazil

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