I have double-checked the closed caps of PCR tubes before placing them in Thermocycler. At the end of the PCR reaction, i set the Thermocycler at 4°C infinity hold. After running the thermocycler, I found the PCR product volume lost/reduced.
Thereafter, I tried another time by covering the PCR tubes' caps with parafilm but after the PCR reaction, the parafilm at the edges of the cap openings had torn (maybe because of heat) then the cap was opened causing a reduction in the PCR volume. Can someone help me with how I can troubleshoot this? what causes this situation?
Do you have a heated lid, or heated lid direct touches your pcr-cap?
if no, this is the reason.
Hi Shalini, Have you tried mineral oil? It's a PCR reagent covering the surface of PCR mixture and reduces the evaporation of reactions run in a Thermocycler without heated lids.
Totally agree with Junjie Shao. If the problem still persist, I would like to proceed your experiment with sealing tape (adhesive plate sealer).
put a (thermal/heating) pad (transfer the heat from lid to cap) between the lid and cap works.
Thermocycler T100, has a lid heating system. Please be sure lid heating is on. If it is on and problem still exist, then please change your PCR tube company. In the picture I see that your tube is melted a bit. During 94 degree steps that might create an evoparation from lid
Hi Shalini...hope one of the above suggestions worked for you. If not, try switching to PCR strips maybe? You also have an option to use "cap-it tool" to help with the domed caps.
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