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Potato Spindle tuber viroids is a low molecular weight, infectious single stranded RNA which is first time discovered in potato. This pathogen has also infected other members of Family Solanaceae. The degradation and instability of any plant RNA is main interruption for dealing with extraction and isolation of these plant pathogenic RNAs and further processing. Mostly RNAs degraded as we take tissues from plants. A very different and brief protocol for PSTVd RNA isolation and storage has been improved and optimized. The present study was aimed to discover and confirm the presence of PSTVd in infected tomatoes in Pakistan. PSTVd infected plants were collected analyzed on the basis of symptoms and signs. The total RNAs was isolated and purified using new optimized Trizol reagent. The RNA of infected tomatoes was converted to cDNA on same day and amplified with reported primers. The identification and confirmation of PSTVd was achieved by BLAST and Sequencing (Accession MK 303578.1. MK977647). The advance optimized Rapid Trizol reagent method was found best and reproducible. The extracted RNA which was stored in Trizol also gave best result even after six months. It is also proved that RNA could be store in trizol reagent for 6 months.
Zaryab Khalid Sial, Farah Khan. (2019) IDENTIFICATION OF POTATO SPINDLE TUBER VIROID (PSTVd) FROM DISEASED PLANTS OF TOMATO (LYCOPERSICON ESCULENTUM MILL.) IN PAKISTAN, , Volume 16, Issue 3.
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