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04-04-2016, 11:51 AM
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#1 |
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Member
Location: cleveland Join Date: Dec 2015
Posts: 26
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16S metagenomics on MiSeq
I will be doing MiSeq run for 16S metagenomics. it will be my first time. Recently some MiSeq v3 300 PE runs gave me trouble and hence I am worried. Are there any things I should do differently from what the 16S protocol says. Illumina asks to spike at 25%, should it be that high.
What other things should be taken into consideration while preparing the libraries. I will have at least 96 samples. Can I pool all on PE 300 v3. Please advise and provide feedback/suggestions, anything and everything there can be for this kind of run. Thanks in advance |
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04-04-2016, 04:13 PM
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#2 |
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Senior Member
Location: California Join Date: Jul 2014
Posts: 198
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There are a ton of threads discussing issues with the 2x300 v3 kits:
http://seqanswers.com/forums/showthread.php?t=59558 http://seqanswers.com/forums/showthread.php?t=67336 |
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04-05-2016, 07:50 AM
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#3 | ||
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Senior Member
Location: USA, Midwest Join Date: May 2008
Posts: 1,178
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Quote:
Quote:
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04-05-2016, 09:43 AM
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#4 |
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Senior Member
Location: CT Join Date: Apr 2015
Posts: 243
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I've had to go to 20% phix for 16S runs otherwise R2 is horrible.
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