Hi, my colleague stored his samples of Drosophila pupae in PCR plate. He just put samples in wells, sealed it with PCR foil and left in room temperature for couple of months. He did not add any silicagel or anything. Now, when we tried to extract DNA with GenAid
Tissue Genomic DNA Mini Kit, we failed in more then half of the samples. Do you have any suggestions how to save the samples and how to extract the DNA?
There are several approaches to recover trace DNA depending on the goal. The general rules are: (1) if the DNA is needed for amplifications only I would advice Direct PCR kits which do not require sample purification. (2) For regular DNA kits, try to concentrate DNA sample by reducing elution volume (vacuumfuge). Even if you are unable to measure the concentration it does not necessarily mean the amplification would not work. Mix reagents first and then add the sample up to the required volume instead of dd water. Try this for the samples you have already extracted with the GenAid kit. (3) If only one PCR is needed (e.q. for genotyping) mix amplification reagents in the tube containing the DNA rather than transferring to a separate tube.
Before Extraction of DNA process start, Dissolve samples in 1X PBS and incubate for Hour, after proper mixing /vortex take volume as per manufacturer instruction using kit. And also follow Anastasia recommendation
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