id	author	title	date	pages	extension	mime	words	sentences	flesch	summary	cache	txt
cord-255194-4i9fc0r7	Djikeng, Appolinaire	Viral genome sequencing by random priming methods	2008-01-07		.txt	text/plain	3776	207	51	An RNase treatment step was added to the SISPA protocol to reduce contaminating exogenous RNAs such as ribosomal RNAs. In the case of polyA-tailed viruses, we perform reverse transcription using a combination of random (FR26RV-N) and poly T tagged (FR40RV-T) primers in order to increase the coverage of the 3' end ( Figure 2 ). Additionally, in order to capture 5' ends of viral RNA, a random hexamer primer tagged with a conserved sequence at the 5' end was added to the Klenow reaction (Figure 2 shows a 5' oligo specific for rhinoviruses). The results of these experiments demonstrate that the SISPA method is very efficient as a genome sequencing method for samples with greater than 10 6 viral particles per RT-PCR reaction ( Figure 5 ). We strongly anticipate that specific adaptations of the SISPA method to conserved regions of different viruses will demonstrate its versatility in a wide range of viral genome sequencing initiatives.	./cache/cord-255194-4i9fc0r7.txt	./txt/cord-255194-4i9fc0r7.txt