M13 helper phage-mediated ssDNA isolation

cw via methods%40net.bio.net (by chengwensu from gmail.com)
Fri Dec 25 12:36:51 EST 2009


Hi,

I have a question about isolating circular ssDNA from plasmids
carrying the f1 phage replication origin - does anyone know if there
is a size limit for the insert that can be subcloned into the MLS? The
problem is I'm working with very big mammalian genomic DNA inserts
(usually 13-16 kb)...and need to grow them in a plasmid with the p15A
ori. Also, does anyone know how to linearize the circular ssDNA
obtained from M13 helper phage-infected bacteria?

Thanks very much in advance for your feedback!


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