Steps for the BSP - (Sep/27/2005 )

Hi everybody,
I'm studying the methylation status of a gene promoter for the first time. I designed the primers for BSP with meth primer, treated my samples with Na-Bis and now I'm going to amplifying my region of interest for sequencing. I have 2 questions:
1) For direct sequencing after amplification can I use the same primers of the amplification itself or am I supposed to use new nested ones?
2) Would it be better to clone the amplified template before sequencing? If yes,why?
Thankyou so much!

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Yes. you can use the same primer for sequencing and use the downstream primer if you are going to sequence only one direction because downstream primer usually yields better result than upstream primer.


Yes. if cost is not a problem, cloning gives better quantitative data.

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