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Forum: Illumina/Solexa 02-14-2018, 02:12 AM
Replies: 3
Views: 2,024
Posted By Justin87
PhiX conc.

Thank you, thermophile!

Sorry for the ignorant question, but just out of interest, could I ask what would happen IF the phix was not spiked into the library at an equimolar concentration and was...
Forum: Illumina/Solexa 02-12-2018, 02:39 AM
Replies: 3
Views: 2,024
Posted By Justin87
16S Sequencing Run - Under-Clustered; Higher PhiX Alignment than expected

Good day,

We recently performed a 16S metagenomics sequencing run (288 samples) on the Illumina MiSeq using a v3 (600 cycle) kit.

We quantified the library using qPCR and calculated the...
Forum: Illumina/Solexa 03-14-2017, 01:26 AM
Replies: 2
Views: 2,565
Posted By Justin87
Reason for POOR per tile sequence quality

We recently performed a sequencing run on the MiSeq sequencer using a V3 (600 cycle kit).

With regards to the per tile quality plot (obtained when using the FastQC analysis software) for one of...
Forum: Illumina/Solexa 01-18-2017, 12:25 AM
Replies: 8
Views: 15,138
Posted By Justin87
Hi All, Appreciate all the feedback! :) ...

Hi All,

Appreciate all the feedback! :)

The DNA had been double-digested. We are performing size selection for an average fragment size of ~500 bp PRIOR to library preparation. Again, I had...
Forum: Illumina/Solexa 01-17-2017, 02:55 AM
Replies: 8
Views: 15,138
Posted By Justin87
Talking Thanks for your response, nucacidhunter! I...

Thanks for your response, nucacidhunter!

I repeated the bio-analyzer run twice, all with very similar results, so that rules out a potentially bad run.

We are pursuing a dd-RAD sequencing...
Forum: Illumina/Solexa 01-17-2017, 01:53 AM
Replies: 8
Views: 15,138
Posted By Justin87
Double Size Selection (500 bp) with AMPure XP Beads

Good day,

Could somebody please elaborate on the ratios of Ampure XP beads required for DOUBLE-SIZE SELECTION to achieve an average fragment size of ~500 bp.

I had followed an illumina...
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