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Old 03-28-2017, 10:35 PM   #1
Location: US

Join Date: Feb 2012
Posts: 19
Question bacterial wgs library loading on MiSeq v3 600 cycles

Just wondering how much wgs library (prepared by Nextera xt kit) should be loaded on MiSeq using v3 600 cycle kit. We tried two times, 20 pM and 14 pM, both runs yielded only ~7 gb (half of 15 gb). We did several amplicon libraries which were pretty good. Are there any tricks? Thank you very much in advance.
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Old 03-28-2017, 11:55 PM   #2
Jafar Jabbari
Location: Melbourne

Join Date: Jan 2013
Posts: 1,234

Your experience indicates issues with correct quantification and loading. For good yield cluster density should be over 1200k/mm2 and this can be achieved with18-19 pM loading.

For specific diagnosis you can contact Tech support at Illumine or post more information about library (trace), quantification method, if bead normalisation was used and run summary page from SAV.
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