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DNA sequence data from both strands was generated from single clones using the primer walking approach, which also conducted with ABI 3730x1 DNA Analyzer.
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A synthetic all-atom model β-sheet consisting of 14 strands was generated by repeatedly excising and aligning strands from the structure of the sucrose-specific porin ScrY from Salmonella typhimurium48.
Various gene-vector directions on two strands are generated by different strand-dependent mutation pressure [16].
Complementary DNA strands were generated from 300 ng mirVana enriched miRNA fractions in a 20 μL reaction volume using the Invitrogen NCode miRNA First-Strand cDNA Synthesis kit.
Then, the second strand was generated to create double-stranded cDNA.
For second-strand cDNA synthesis, the mRNA template was removed and a replacement strand was generated to form double-stranded (ds) cDNA.
The second strand was generated using a SuperScript Double-Stranded cDNA Synthesis kit (Invitrogen, Camarillo, CA), purified via magnetic beads, the ends repaired and a single adenine base added to the 3′ ends.
Briefly, a second DNA strand was generated.
The first cDNA strand was generated using RevertAid™ First Strand cDNA Synthesis Kit (Fermentas, Thermo SCIENTIFIC, USA) following the manufacture' protocol.
The first cDNA strand was generated using a Takara Reverse Transcription System (Japan) following the manufacturer's protocol.
The first cDNA strand was generated using the Takara Reverse Transcription System (Japan) by following the manufacturer's protocol.
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