DNA size selection method for long read sequencing
Technology
An LRS method that allows simple selection of fragments larger than 7.5 kb in length from a fragmented nucleic acid sample using paramagnetic Solid Phase Reversible Immobilization (SPRI) beads in a binding buffer.
The protocol accommodates the desire for a technology for size selection >10kb that is tolerant to a wide range of input amounts, does not add complexity to laboratory workflows, is amenable to automation and reliably recovers high yields of on-target library fragments.
Advantages
- Simple: modifies one step of the current protocol without major adjustment to liquid handling steps, ideal for automation.
- Sensitive: allows removal of fragments smaller than 7.5 kb and is also tolerant to low inputs (i.e. <10ng/μL).
- Cost-effective: does not require the use of expensive reagents.
- Validated: on PacBio sequencers and protocols.
- Flexible: tolerant to a wide range of input amounts.
Applications/Context
LRS and any protocols where size selection of the DNA fragments requires removal of fragments smaller than 7.5kb. Our method has already been optimised for PacBio protocols by replacing one step of the current workflow.
*Header image credit: Andy Leppard, Wikimedia
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