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The protocol for cDNA library generation we describe is independent of locus specific amplification primers and can be used for sequencing any unknown type of influenza A viruses.
In case of single restriction enzyme digestion, both anchors can potentially ligate to both MseI generated sticky end but only AB or BA containing fragments can be used for sequencing.
We conclude that DNA extracted from FFPE tissues can be used for Sequencing by Synthesis (SBS) and that neither an increase in ischemia time from 20 min. to over 360 min. nor a longer fixation time of up to 72 hours plays a major role in sequencing quality (Figure 1, Table 1).
In fact, it has been hypothesized that this configuration could be used for sequencing DNA with single-base resolution.
An M13 sequence tag was added to the 5′ end of each primer to be used for sequencing.
These multi-gene families can be used for sequencing error detection, mutation identification, and pangenomic profile computation [19].
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PCR primers were used for sequencing.
Both forward and reverse primers were used for sequencing.
Both sense and antisense inner primers were used for sequencing.
Primers P1, P2, or P38 (5'AGGCTTGTGCAGGCAGTG3') (68) were used for sequencing reactions.
Two µl of the unpurified PCR product was used for sequencing both strands of the product.
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