We stiIl use pestle and mortar which is very time-consuming and we would like to speed up this process.
We've had similar machine (Retsch) - I think Qiagen is just a rebranded model. I have found using it quite tedious as the jars are very expensive and had to be sterilized between different tissues. There are adapters for 1-2 ml tubes ( about $1200) but we needed to grind at least 0.5 g of tissue and for that we need tubes more than 5 ml capacity. We use now Interscience's MiniMix or Stomacher. The grinding in plastic bags is gentle so temperature is not a big factor. This method works for majority of our isolations. If you need to grind more challenging tissues you can use any bead beater - like a Qiagen machine - but the ones which use disposable sterile containers - for example Precellys. There is even machine which uses regular 1.5 ml tubes up to 14 and 15 ml (Next Advance or Mo-Bio) - There is a lot of them right now on the market.
I used that instrument commonly for lung virus titers of a gammaherpes virus, as well as occasionally other tissues (spleen, livers) to look for reactivated virus. Worked well. We did a side by side comparison and found equivalent viral titers in a plaque assay. We used the disposable tubes...we found you can wash and autoclave the metal beads and reuse them (again, tested to make sure they were virus free afterwards). Did not reuse tubes (disposable, plastic, I believe 2 mL size)We then transferred the tissue that was in 1 ml of media to a larger tube to make serial dilutions for the assay.
Hi Claire - Could you kindly let me know what speed and time you use for your isolation? I am trying to use it for isolating RSV from lungs to determine viral titres by plaque assay.
Hello Marsha - I would suggest checking out Precellys website http://goo.gl/c17OZC
Both Precellys and Qiagen work with beads of various sizes and composition. The protocols would be similar. Definitely you don't want to overheat the sample.
I asked Precellys similar question and they suggested 2-3 cycles 40 second each with CK28 beads (ceramic 2.8 mm). I am sure you can buy equivalent beads at vwr or Fisher. For Qiagen we were getting the beads from Biospec www.biospec.com. They also have nice info about beads http://goo.gl/Yy7ZVf
happy grinding!
my samples are in 50mL disposable tubes, I want to avoid to transfer them to new tubes. i have a hand-made die for the tubes, but the plastic tube break up inside it due to physical force of the beads.......any suggestions.
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