Hi there,
I am supposed to amplify some promoters cloned in pBACe3.6 vectors. I am fairly new with this system or promoter cloning, I was thinking if any experts put there could help me? My first question is, can I use BAC vectors directly and PCR template or do I need to digest it first and use the digested product for PCR template? Any help or suggestion is highly appreciated.
many thanks in advance.
You can use the BAC vector as a template directly for PCR amplification of promoter of a gene cloned into that vector. No need for digestion! But primer design must be very gene-specific.
Thanks Asim. I will try that. The primers are very specific to that promoter region.
Cheers.
Additionally while performing experiment try to use the different concentration of template (BAC vector|). This way you will get the idea at which concentration you are getting maximum amplification.
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