id	author	title	date	pages	extension	mime	words	sentences	flesch	summary	cache	txt
cord-255043-uxdsjr39	Bustin, Stephen A.	RT-qPCR Testing of SARS-CoV-2: A Primer	2020-04-24		.txt	text/plain	4552	201	46	The only reliable test in current use for testing acute infection targets the genome of SARS-CoV-2, and the most widely used method is quantitative fluorescence-based reverse transcription polymerase chain reaction (RT-qPCR). Given that on 9th January 2020 SARS-CoV-2 was definitively identified by the Chinese CDC as the causative agent for COVID-19 pneumonia and that its genomic sequence (GenBank accession number MN908947) was made available on 10th January, it is extraordinary that by the time the earliest documented transmission within the UK appeared on 28th February, no definitive action plan, stockpile of assays and required consumables, RNA extraction robots or high throughput qPCR instruments had been assembled to allow immediate and widespread RT-qPCR testing. This involves designing assays that generate PCR products of 60-80 bp, using fast RNA-and DNA-dependent DNA polymerases such as Superscript IV and KAPA Taq polymerase, respectively, and selecting instruments capable of rapid cycling, for example Eco from PCRMax. This allows the RT step to be limited to 2 min or less, with the denaturation and annealing/polymerisation steps limited to 1 s each ( Figure 3) .	./cache/cord-255043-uxdsjr39.txt	./txt/cord-255043-uxdsjr39.txt