Synthesizing the plasmid

Influenza A virus, Udorn human strain of influenza A virus , genome was converted to cDNA by reverse transcription RNA is made into DNA using a DNA polymerase . The M gene on the cDNA was amplified by polymerase chain reaction (PCR) with primers that were specific for the M gene. Point mutations at amino acid # 27(M2V26T), #30(M2A30P), and #31(M2S31N) and a deletion of amino acids 29-31 (M2del29-31) were amplified. The product of the cDNA amplification (construct) was cloned into a vector (now called a plasmid) and used to generate mutants through reverse transcriptase PCR PCR in which the starting template is RNA, implying the need for an initial reverse transcriptase step to make a DNA template. .