I need to now how can we make DNA extraction from hard ticks? I will be grateful for help
Hi Nuha, you could try cutting or bead beating techniques, or if you require the DNA to be intact try a buffer such as MicroLysis Plus from Microzone and mix with the cells before thermocycling.
Hi Nuha, one other thing you could try is grinding under liquid nitrogen if you have access to it as well as a mortar and pestle. A beadmill may work out better for you though.
I successfully extracted gDNA from insects (mosquitoes) using DNAzol. I normally squish insects in few microliters of DNAzol (20 to 100 depending upon size and numbers) in a 1.5 mL vial using a sterile hard plastic pestle. Then I wash the pestle with additional microliters of DNAzol prior to extraction. After that, the protocol itself is pretty straightforward and quick but in case you're going to try and you need any further info feel free to ask.
P.s. It's not toxic as organic reagents are and obviously less dangerous. The cost also is lower if compared.
In addition, after you use lysis buffers (cell or nuclei) avaoid vortexing. I also used liquid nitrogen for grinding as well as 26 gauge needle with syringe for breaking the cell membranes.
Summary: Look for the best kit you want to use for gDNA extraction from ticks, then grind the ticks and store them. At the time of extraction, follow the protocol and just break the tissue by grinding using liquid nitrogen.
I recommend Alex suggestion, one other thing you could try is grinding under liquid nitrogen if you have access to it as well as a mortar and pestle. A beadmill may work out better for you though.
DNA extraction. Available from: https://www.researchgate.net/post/DNA_extraction6 [accessed Aug 15, 2017].
Thanks for all answers and help. If anyone have the protocol of the work I will be grateful to him and any pdf or papper to help me because I need it urgently. Thanks again.
To the protocol mentioned by Dr. Paolo Scarpelli, you can add a step of freeze thawing before squish insects as it will help in making the insects soft for squishing.
I did this with Aedes aegypti and worked well. Thanks
Hi Nuha,
try this...
https://www.vetres.org/articles/vetres/pdf/2004/06/V4025.pdf
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