View Single Post
Old 01-06-2017, 06:41 AM   #5
gringer
David Eccles (gringer)
 
Location: Wellington, New Zealand

Join Date: May 2011
Posts: 823
Default

You assemble a large region by having multiple DNA copies. Consider a situation where DNA is extracted from a colony of 10,000 bacteria. In an ideal world there would be 10,000 copies of the bacterial DNA floating around in that extract, even without any amplification. Practically, the amount will be considerably less than this, but the recommended protocols use far in excess of this coverage amount.

The standard MinION sample prep requires a reasonable amount of DNA to be present (but not out of the ordinary for high-throughput sequencing): about 200ng for the rapid sequencing kits. This represents an amount on the order of 33k unamplified copies of DNA for human-sized genomes, so there shouldn't be a problem with getting 1000 copies of sequence to cover a particular region, especially if it's bacterial DNA.

Last edited by gringer; 01-06-2017 at 05:50 PM. Reason: updated according to AllSeq's calculations
gringer is offline   Reply With Quote