The simple method of nucleic acids extraction, based on guanidine thiocyanate extraction buffer, was sufficient for obtaining a good templates for RT-PCR. The RT-PCR reactions were performer from the start to the end (both the reverse transcription and the HLVd-cDNA amplification) in the same reaction mixture and in the very small volume of reaction (10 μI). Both pairs of primers designed by authors were good for reverse transcription and later for amplification of the HLVd-cDNA. The presence of gelatin as a stabilizer of DNA polymerase was indispensable for successful performance of RT-PCR.
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