Reagents to complete depletion/size selection of DNA <25 kb
The SRE kit uses size-selective precipitation to dramatically reduce short reads below 25 kb.
Short DNA fragments <25 kb are progressively depleted (i.e., depletion efficiency increases as DNA gets shorter). DNA <10 kb are nearly completely removed. Depletion range estimated based on sequencing read length distributions and gel electrophoresis.
The read length enhancement is dependent on:
1. the amount of short DNA present in the input DNA
2. the amount of long DNA present in the input DNA
Read length N50 is increased by up to 25 kb on Oxford Nanopore MinION/GridION/PromethION.
PacBio Sequel applications are coming soon.
Overall recovery efficiency is dependent on:
1. the quality of the input DNA
2. the input DNA concentration.
Expect to recover 50–70% of your HMW DNA.
Sample must be predominantly HMW DNA (>48 kb) or recovery efficiency will be compromised.
Processing requires only a simple centrifuge step and two wash steps. Total process time is one hour.
1. Buffer SRE — 1 mL
2. Buffer EB — 5 mL
Sufficient for at least 16 standard purifications.