<p>The performance of each sequencing primer is described in <a href="http://www.pl... more <p>The performance of each sequencing primer is described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0064785#pone-0064785-t001" target="_blank">Table 1</a>, as seen by the average percentage of bases generated from the 30 complete genomes with QV more than 30 and 40, respectively. The QV values were generated using the proprietary sequencing analysis software (version 5.2) of the ABI 3130×l genetic analyzer (Applied Biosystems). Length of Read (LOR) is defined as the length of sequence with QV20 and above for at least 20 continuous bases.</p
<p>The white and black columns represent the percentages of D151 mutations in MDCK-cultured... more <p>The white and black columns represent the percentages of D151 mutations in MDCK-cultured isolates deposited in GISAID EpiFlu database (n=2729, 2004-2012) and from this study (n=77, 2009-2011), respectively. The relative trend in an increasing proportion of in-house MDCK-cultured isolates collected from 2009-2011 is similar to that for the GISAID sequences.</p
<p>The T<sub>aS</sub> for all the PCR primers ranged between 58 and 61°C. MBTun... more <p>The T<sub>aS</sub> for all the PCR primers ranged between 58 and 61°C. MBTuni-12 and MBTuni-13 primers targeting the 5′ and 3′ ends of each segment were adopted from published methods <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0064785#pone.0064785-Chan1" target="_blank">[21]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0064785#pone.0064785-Zhou1" target="_blank">[27]</a>, with nucleotides (in bold) representing the modifications made. Nucleotide R (bold) in the primer sequence indicates a degenerate nucleotide that represents A or G.</p
<p>The performance of each sequencing primer is described in <a href="http://www.pl... more <p>The performance of each sequencing primer is described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0064785#pone-0064785-t001" target="_blank">Table 1</a>, as seen by the average percentage of bases generated from the 30 complete genomes with QV more than 30 and 40, respectively. The QV values were generated using the proprietary sequencing analysis software (version 5.2) of the ABI 3130×l genetic analyzer (Applied Biosystems). Length of Read (LOR) is defined as the length of sequence with QV20 and above for at least 20 continuous bases.</p
<p>The white and black columns represent the percentages of D151 mutations in MDCK-cultured... more <p>The white and black columns represent the percentages of D151 mutations in MDCK-cultured isolates deposited in GISAID EpiFlu database (n=2729, 2004-2012) and from this study (n=77, 2009-2011), respectively. The relative trend in an increasing proportion of in-house MDCK-cultured isolates collected from 2009-2011 is similar to that for the GISAID sequences.</p
<p>The T<sub>aS</sub> for all the PCR primers ranged between 58 and 61°C. MBTun... more <p>The T<sub>aS</sub> for all the PCR primers ranged between 58 and 61°C. MBTuni-12 and MBTuni-13 primers targeting the 5′ and 3′ ends of each segment were adopted from published methods <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0064785#pone.0064785-Chan1" target="_blank">[21]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0064785#pone.0064785-Zhou1" target="_blank">[27]</a>, with nucleotides (in bold) representing the modifications made. Nucleotide R (bold) in the primer sequence indicates a degenerate nucleotide that represents A or G.</p
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