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Old 05-10-2019, 04:00 AM   #6
kmcarr
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Location: USA, Midwest

Join Date: May 2008
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Quote:
Originally Posted by SDPA_Pet View Post
Thanks. I just want to make sure if the sequencing primers for bridge amplification step for Illumina sequencing? Am I right? I just want to know which step the sequencing center would use these sequencing center.

When I check the youtube video about illumina sequencing, I don't think illumina requires a sequencing primers to sequencing.
Yes, Illumina sequencers absolutely require sequencing and index read* primers. Bridge amplification and sequence generation are separate things. Every Illumina reagent kit/cartridge has tubes/wells of dedicated read 1 (R1), read 2 (R2) and index 1 (I1) primer mixes. Your custom primers will be mixed into those. Illumina provides specific guidelines for the concentration of custom primers to mix in. Make sure that your sequencing provider is familiar with adding custom primers to a reagent kit/cartridge.

*The exception to separate primers is for index read 2 (I2) in dual index situations. That read is primed using the oligos grafted to the flow cell surface. That's why the dual index 16S-V4 protocol includes custom primers for R1, R2 and I1 but not I2
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