What about extraction from cell lines with no connective tissue etc. Would it be sensible to decrease the 30 minute extraction step in case the genomic DNA becomes badly degraded?
Hey Ian, that's a great question. I don't know the answer to this right now, but you might find more info in the published literature. What specific sample type do you have in mind? As an example, HotSHOT has been used effectively for e.g. saliva. My initial gut response is that I wouldn't anticipate the temperature or time in the protocol to have a substantial impact. Is there a reason you are concerned about non-connective tissue? From my experience, degradation is usually caused by chemical reactions (e.g. DNAases) over time. Those are some initial thoughts - I hope it's helpful! If you are planning to do an experiment, keep us in the loop! We'd love to share the results. Are you working on a project where you are considering the HotSHOT method? Have you had a look at the Dipstick method as well? ua-cam.com/video/i5JmZ8n13es/v-deo.html
What about extraction from cell lines with no connective tissue etc. Would it be sensible to decrease the 30 minute extraction step in case the genomic DNA becomes badly degraded?
Hey Ian, that's a great question.
I don't know the answer to this right now, but you might find more info in the published literature. What specific sample type do you have in mind? As an example, HotSHOT has been used effectively for e.g. saliva.
My initial gut response is that I wouldn't anticipate the temperature or time in the protocol to have a substantial impact. Is there a reason you are concerned about non-connective tissue? From my experience, degradation is usually caused by chemical reactions (e.g. DNAases) over time. Those are some initial thoughts - I hope it's helpful!
If you are planning to do an experiment, keep us in the loop! We'd love to share the results. Are you working on a project where you are considering the HotSHOT method?
Have you had a look at the Dipstick method as well? ua-cam.com/video/i5JmZ8n13es/v-deo.html
Very simple and versatile protocol, great product. Was it tested on human saliva yet ? (prior to PCR)
Thanks! There are definitely people who have used it to extract DNA from saliva (for PCR).