Demonstrated Protocol, Last Modified on January 12, 2018, Permalink
The Chromium™ Genome Protocols generate long-range information across the length of individual DNA molecules. Starting the process with High Molecular Weight (HMW) genomic DNA (gDNA) will typically result in better application performance, such as increased haplotype phase block length and ability to increase sensitivity for large structural variants. Optimal performance has been characterized on input gDNA with a mean length >50 kb.
This Demonstrated Protocol outlines a method for gDNA extraction from a single Drosophila melanogaster larva, a single Drosophila melanogaster adult, and a single Aedes aegypti pupa. This Protocol can produce gDNA from ~48.5 kb – >200 kb from a single insect as analyzed on a pulsed-field gel, and typically >40 kb after the Chromium Genome Protocols. There are many variables that will affect gDNA quality in sample types, including sample age, transportation methods, type of sample, additives, freezing method, etc.
While this Protocol is demonstrated with Drosophila melanogaster and Aedes aegypti, it may be used for gDNA extraction from other types of insects. Modifications to this Protocol may be necessary for other sample types.