I ran a PacBio sequencing run with BstEII digested lambda DNA.
https://www.neb.com/products/n3014-d...%20Information
Since the DNA is digested with a single restriction enzyme, there should be equal moles of each fragment. However, in the run I saw few read counts at the smaller fragment size, then increasing as size got larger, and a slight drop off for the largest fragments.
One of the larger fragments was also a significant outlier, having 1/3 the counts as the next size up and down. This is concerning because it may mean there is some sizing bias in the run. What could be causing this?
https://www.neb.com/products/n3014-d...%20Information
Since the DNA is digested with a single restriction enzyme, there should be equal moles of each fragment. However, in the run I saw few read counts at the smaller fragment size, then increasing as size got larger, and a slight drop off for the largest fragments.
One of the larger fragments was also a significant outlier, having 1/3 the counts as the next size up and down. This is concerning because it may mean there is some sizing bias in the run. What could be causing this?
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