How to clean-up 96-well microplates? (View forum version)



phucvn

Posted 09 June 2010 - 07:35 PM

Hi All,

I am in a poor lab and we are in short of budget. I am thinking a way to clean 96-well microplates to re-use in normal PCR such as insert check, or even in sequencing. I tried once with multiple washes by ultrasonication in dDW and UV but I am not sure if it worked. I searched out some kinds of 96-well plate washers but it is impossible for us to buy now. So I would like to ask you if anyone has experienced in this question.
Do you have any other suggestions?

Thanks

perneseblue

Posted 09 June 2010 - 08:00 PM

Here is my suggestion.

Take your plate and soak it in beaker of warm 1M HCl and leave the PCR plate to soak overnight.
The HCl should hydrolyse all DNA.

After take your plate and wash out with sterile miliQ water. Then dry in 50C incubator. If you have access to UV, you might want to expose the dried plate to a good dose of UV to denature any DNA.

you might want to give this idea of a protocol a test. Run a PCR in the plate. Then clean up the plate. Then run the same PCR but without the template. If you get no PCR product on the reused plate, you are good to go.

moljul

Posted 10 June 2010 - 06:47 AM

Here is my suggestion.

Take your plate and soak it in beaker of warm 1M HCl and leave the PCR plate to soak overnight.
The HCl should hydrolyse all DNA.

After take your plate and wash out with sterile miliQ water. Then dry in 50C incubator. If you have access to UV, you might want to expose the dried plate to a good dose of UV to denature any DNA.

you might want to give this idea of a protocol a test. Run a PCR in the plate. Then clean up the plate. Then run the same PCR but without the template. If you get no PCR product on the reused plate, you are good to go.


what about nucleases?? i would be afraid of contaminating my plates with e.g. RNases!

phage434

Posted 12 June 2010 - 12:50 AM

I agree that this is a good protocol. You might want to neutralize the HCl with Tris and multiple washes of DI water. I doubt that there is any necessity for the UV. RNAses will not be removed, but that's ok, since you usually add RNAses to your DNA. If you are doing RNA work, then you have another set of problems, and I would not recommend reusing plates.

phucvn

Posted 16 June 2010 - 02:14 AM

Hello Perneseblue, Veteran, and Phage343,

Thank you so much for your suggestions and ideas. I will try and repost the results. Surely that for RNA assays, I must be careful and carry out with brand new labwares.
Cheers,