Next generation sequencing
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23 months ago

I am studying about NGS for my exams. I have a doubt regarding the labelling(not scientific Labelling) strands as forward and reverse strands in NGS. Anyone care to explain?!

dna sequencing Strand Ngs • 698 views
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Sequencing is always in 5' --> 3' direction. Generally reference genomes include a strand that is published as "top" strand (that data goes in 5' --> 3' direction). When you align your NGS data the aligner will automatically align the data to both strands and included that information in alignment file.

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23 months ago
Trivas ★ 1.7k

The strands aren't Forward and Reverse for NGS; they are called R1 and R2 (read 1 or read 2) depending on if they have the P5 or P7 sequencing adaptor on the 5'-end respectively.

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23 months ago
JustinZhang ▴ 30

If you mean R1 and R2, here are representative intro videos for you:

Illumina sequencing (Brige): https://www.youtube.com/watch?v=fCd6B5HRaZ8

MGI sequencing (Ball): https://www.youtube.com/watch?v=RGcpftDHpng


And, for what's the practical significance of R1 and R2, here are one discussion:

GATK forum: https://gatk.broadinstitute.org/hc/en-us/community/posts/360075017111-strand-bias-and-orientation-bias

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