Hot StarTaq Master Mix Kit (2500 U)

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Protocol tips
Follow manufacturer's instructions

Publication protocol

HCV RNA was extracted from 140 μl plasma using a commercially available kit (QIAmp viral RNA kit; Qiagen, Tokyo, Japan). To amplify the NS5B region of the HCV genome, the extracted RNA was reverse transcribed and amplified using SuperScript One-Step reverse transcription-PCR (RT-PCR) (Invitrogen, Tokyo, Japan) and a set of primers. PCR amplifications using outer primers (nucleotides [nt] 7999 to 8825) and inner primers (8159 to 8630) were performed as previously described (1) using Hot Star Taq master mix (Qiagen). The amplified fragments were sequenced by a direct sequencing method with the BigDye Terminator v1.1 cycle sequencing kit and an ABI Prism 310 sequencer (Applied Biosystems, Foster City, CA, USA). Based on the sequence similarity to the reported sequences from the international DNA databases (DDBJ, EMBL, and GenBank) using the program Genetyx-Win version 9.0 (Genetyx Corporation, Tokyo, Japan), each HCV isolate was assigned an HCV subtype (2). The HCV genotypes/subtypes were reexamined to confirm the HCV genotype/subtype data obtained from the medical records.

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Manufacturer protocol

Download the product protocol from Qiagen for Hot StarTaq Master Mix Kit (2500 U) below.

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