Briefly, I need a cost and time effective method - which completely exludes the possibility of contamination between samples - in order to get ranaviral DNA from the liver of EtOH stored tadpoles.
Criterions are the following:
- For DNA extraction I will use Wizard® Genomic DNA Purification Kit Protocol (Promega).
- I don't have bead beater or tissue lyser machine just an Eppendorf thermomixer.
- Large sample size (approx. 500 samples).
Many thanks in advance.
Dear Traci L Testerman ,
Many thanks for your effort. Maybe a vortex combined with stainless steels bead (2.8 mm diameter) will give the required homogenity. I will definitely give a try for that.
Cheers,
David
Hi,
I have a practical solution for homogenising soft tissues. A rotary tool called Dremel is very handy and budget solution. There are thousands of bits available that can be used for homogenising tissues. Most of hand on tissue homogenisers are based on Dremel tool. If you obtain a Dremel tool, I suggest this type of bits.
Good luck
Link: https://encrypted-tbn0.gstatic.com/images?q=tbn%3AANd9GcRLrlEfo9A32jhade3swsAHfGQcILK0sGGQtGkTO_nGYJEiYjfC
Hey Kaan Hürkan ,
Many thanks for this idea. I have two problems with dremmel. I am not sure about that the smallest bit fits to an 1.5mL eppendorf vial and If it fits there is the issue of contamination. I have a relatively large sample size (N=500) and short time for desinfection between samples. This methods wouldn't be so practical in my situation but It could be nice in the case of a few samples.
Cheers,
David
Hi dear Dávid Herczeg how arre you doing?
This is the smallest bit of Dremel (https://www.cthardware.com/tools-68/powertools/special-tools/special-tools-dremel-accessories/dremel-108-1-32-0-8mm-engraving-cutters.html).
They are autoclavable since being metal.
Cheers to everyone in Debrecen!
I am doing great Kaan Hürkan hope you are doing fine as well. :)
Cheers,
David
The tip of the bit can be far away from the Dremel tool's chuck that holds it. Maybe even 4 or 5 cm. This allows you to "buzz" the tip and long shaft in disinfectant and spin it dry- as in a small hot air blast. Check the system to see how long to buzz and heat...for higher speed sampling.
With what you have available, you may try to space yourself if you do come back to doing it manually. Patience is key. Also manual homogenizing will best prevent any instance of contamination. I believe you have disposable pestles which saves you the problem of having to disinfect each time you use one. I hope this helps if it becomes necessary.
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