Hey lab folks, for PCR do you prefer disposable (plastic or metal, one time use like https://www.fishersci.com/shop/products/corning-sealing-mat-384-well-polypropylene-storage-plates/07202500) or reusable (usually silicone, washable and autoclavable like https://www.fishersci.com/shop/products/axygen-impermamat-axymat-sealing-mat-2ml-96-well-deep-well-plates/14222010) type plate sealers? Which would reduce contamination the most? I will be doing microsatellites for the most part.
I use reusable plate sealing mat for my PCR. I have not faced contamination problem as far, even after reusing the mat multiple times.
I would recommend reusable plate sealer but care should be taken while washing and autoclaving to avoid contamination.
I use reusable plate sealing mat for my PCR. I have not faced contamination problem as far, even after reusing the mat multiple times.
I would recommend reusable plate sealer but care should be taken while washing and autoclaving to avoid contamination.
This is an interesting question. We use strip caps but has anyone ever tried coating the caps with a mineral oil? or something similar?
I like using silicone mats for PCR because they stay on so well and are reusable. Like Kiruthika, I didn't have any issues with contamination with that approach. At Northern Arizona University, they let them soak, scrub them, rinse with DI water, and air-dry for 24 hours. No issues so far for several years.
For storage, I like foil plate seals, because they are quick to get on and you can puncture the top with pipet tips if you want to access particular wells. Then you can cut strips of the seal to re-seal those wells, or use an entire new one to seal the whole plate.
Hi Sean, I have used for many years the reusable ones, mainly for microsats and Sanger sequencing. I never had a problem with contamination (even in a quite busy lab). We used to wash, leave on a bleach solution, rinse with water, and every now and then autoclave.
I would definitely not recommend if you are doing deep sequencing (coverage >1,000), in this case always use disposable materials and tips with filter.
Thanks everyone! Thus far I have used reusable ones as well. Sounds like I'll keep it up. But I like the idea of foil for storage Karl- good thoughts. Thanks!
For AFLP and microsats i used to use silicone mats. I didn't have any issues with contamination with that approach. After use i store them in mq water till container is full. Then i rince them with 0.5 M NaOH (which i re-use many times) and rince them 5 times with Demi. I dry them and store them in old pipet tip boxes (a box for intact 96 mats, a box for half mats and a box for smaller peaces of mats...
no contamination issues at all...
You can use te mats for years...
For storage, I also like foil plate seals, because they are quick to get on and you can puncture the top with pipet tips if you want to access particular wells. Then you can cut strips of the seal to re-seal those wells, or use an entire new one to seal the whole plate. BE aware of glue accidentally sealing of pipet tips when pearchin alu foil.. always check what you pipet...
Greetz,
niels
What is the risk of contaminating samples with the adhesive seal if you are say shipping plates to an external lab i.e., if the plate gets turned upside down and the sample is in contact with the foil?
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