3:37 You only mentioned ethanol here. The protocol I have suggests an ethanol(90ml)-AL(86ml) mixture. Thanks for the video though. For someone like me who never worked in a lab, it was very useful.
@beckycloud8308
10 ай бұрын
My lab is worried about splash risk while working with field-collected mosquitoes that may contain human pathogens, so pestle homogenization is not ideal for me. Would 3mm stainless steel beads work fine?
@itsjatt322
Жыл бұрын
i have one question. can we preserve whitefly in 70% Ethanol and extract DNA after 2 months? is it possible?
@juanayala7767
3 жыл бұрын
Thanks for video! It has helped me a lot. I just have one question. I noticed that in the DNA precipitation step, you didn't use the AL buffer as is described in the manual of the product. So, is that buffer optional or just for specific samples?
@WolbachiaProj
3 жыл бұрын
Great question. Buffer AL is categorized as a lysis buffer, so we added it at 2:47 during the "Cell Lysis and DNA Precipitation" portion of our protocol. Here's a link to the protocol we use, which uses the DNeasy kit for insect extraction: www.qiagen.com/us/resources/resourcedetail?id=cabd47a4-cb5a-4327-b10d-d90b8542421e&lang=en I hope that helps, please let us know if you have further questions! Also feel free to email us at wolbachiaproject(at)vanderbilt(dot)edu.
@tasnimbintaosman590
Жыл бұрын
We Didn’t get dna. All lines we followed except 200 protinase K. We used 30 protinase K. Among 8 samples, we got for two.
@WolbachiaProj
Жыл бұрын
This protocol is optimized with a shorter incubation time for classroom use. If you reduce the amount of proteinase K, we recommend increasing the incubation time to enhance lysis. Also make sure to efficiently grind the sample with micropestles.
@arumoe
7 ай бұрын
Thankyou so much ❤️
@anastasiating1834
Жыл бұрын
Can we extract the DNA from the legs of insects? And do we need to get a fresh sample or alive insects for DNA extraction
@saranga6304
Жыл бұрын
Romario, its good that u also watched this video.😄
@anastasiating1834
Жыл бұрын
@@saranga6304 🤣🤣🤣🤣
@WolbachiaProj
Жыл бұрын
Yes, Wolbachia can be detected in legs as reported here: www.ncbi.nlm.nih.gov/pmc/articles/PMC3342236/. Most of our participants isolate from fresh samples, but some have successfully amplified DNA from from dead organisms. Make sure to indicate the state of your organism along with any reported data.
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