I would like to obtain high-quality DNA. There are a lot of protocols for DNA isolation but most of them are for PCR purpose and they are not appropriate for sequencing due to low quality.
Thanks in advance for tips!
For high recovery of RNA from yeast, my lab uses a hot phenol extraction. The protocol we use is from Kohrer and Domdey, 1991, methods Enzymol, 194: 398-405.The procedure is very dangerous and requires working in a hood. The benefits outweigh the risks because this is the gold standard for RNA isolation. I attached our protocol.
For DNA you would just have to modify the pH of the phenol to select for DNA instead of RNA.
I'm a fan of the MoBio kits. Easy to use and good quality DNA without having to mess around with toxic chemicals such phenol/chloroform
I use a mix-protocol with both mechanical and chemical action on the yeast cell wall, with glass beads and guanidine thiocyanate followed by a standard DNA extraction procedure. Has worked well even with difficult yeast such as Malassezia.
Have you suggestions to isolate DNA from insects (aldo for genome sequencing)?
I personally eventually used kit from EurX: Bacterial & Yeast Genomic DNA Purification Kit and extracted DNA worked perfectly for sequencing :)
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