Reagents to complete depletion/size selection of DNA <25 kb
The SRE kit uses size-selective precipitation to dramatically reduce short reads below 10 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
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 (>30 kb) or recovery efficiency will be compromised.
Processing requires only a simple centrifuge step and two wash steps. Total process time is two hours with 10 min hands on time.
1. Buffer SRE — 1 mL
2. Buffer EB — 5 mL
Sufficient for at least 16 standard purifications.