DNA EXTRACTION
Most commercial kits use silica-based filters but a wide variety of options are available. During the workshop we follow a simple alkaline lysis method of DNA extraction (Jisming-See et al. 2016 adapted from Ivanova et al. 2009). Other extraction options are presented in the DNA barcodes for insects book chapter.
The DNA Bank network conducted an informative comparison of different DNA extraction products.
The DNA Bank network conducted an informative comparison of different DNA extraction products.
As always, before beginning you lab work make sure to wear clean gloves, clean the bench and work over a KimWipe.
ALKALINE LYSIS DNA EXTRACTION
a. Add 17.5ul alkaline buffer to each tube.
b. Incubate the tubes at 95C for 20 minutes.
c. Add 32.5ul neutralization buffer to each tube.
The image below from the Diamantina Institute show the rationale behind the alkali lysis extraction method.
b. Incubate the tubes at 95C for 20 minutes.
c. Add 32.5ul neutralization buffer to each tube.
The image below from the Diamantina Institute show the rationale behind the alkali lysis extraction method.