Second, the T-vector cloning strategy was employed to generate the recombinant plasmid for direct sequencing with an ABI Sequencer 3730.
All vectors were verified by sequencing with an ABIPrism 3000 sequencer.
The identity of the products was verified by sequencing, with an automated DNA sequencer.
All cloned sequences were confirmed by DNA sequencing with an ABI 3100 automatic sequencer (PE Applied Biosystems, Foster city, CA).
Correct sequences were verified by double-stranded DNA sequencing with an ABI PRISM 310 Sequencer (Applied Biosystems) prior to expression and purification.
Sequence-specific walking primers were designed for both strands of each fragment and were employed for sequencing with an ABI 3730 DNA sequencer (Applied Biosystems, USA).
Following quality assessment, poly (A) mRNA preparation and sequencing with an Illumina HiSeq 1000 sequencer (Illumina Inc., San Diego, CA, USA) were performed as described previously [ 52].
Only samples in which rhinovirus had been identified by real-time PCR were submitted to 5′ ΝCR sequencing with an ABI 377 automated sequencer (Applied Biosystems), and results were submitted to the GenBank database, accession nos.
