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Table 1 Primers used for amplification of PB2 and PB1 gene segments and subsequent cloning into pHW2000 vector

From: A ligation and restriction enzyme independent cloning technique: an alternative to conventional methods for cloning hard-to-clone gene segments in the influenza reverse genetics system

Gene

Forward Primer (5′ to 3′)

Reverse Primer (5′ to 3′)

Expected Size (bp)

PB2

TCCGAAGTTGGGGGGGAGCGAAAGCAGGTC

CCGCCGGGTTATTAGTAGAAACAAGGTCGTTT

2370

PB1

TCCGAAGTTGGGGGGGAGCGAAAGCAGGCAAAC

CCGCCGGGTTATTAGTAGAAACAAGGCATTT

2370

  1. The forward and reverse primers contain 16 and 13 nucleotides, respectively which are complementary to the pHW2000 (italics), followed by 12 and 13 nucleotides forming conserved UTR (bold) and 2–6 gene specific nucleotides (underlined). The nucleotide sequences (in italics) anneal to the pHW2000 and allow directional cloning of desired gene segment into the pHW2000 vector
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Virology Journal

ISSN: 1743-422X