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Old 12-20-2011, 08:13 AM   #1
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Location: Cambridge, MA

Join Date: Aug 2010
Posts: 6
Default Ion Sphere ID summary, polyclonal %

For our first runs we have "Filtered: Polyclonal" in the 25-35% range. Is this due to more beads put into the emulsions than necessary, non-accurate library titration or possibly >1 bead in a well? Is this percentage in the expected range?

Last edited by The_Dark_Base; 12-20-2011 at 03:40 PM.
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Old 12-21-2011, 01:53 AM   #2
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Location: Australia

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Hi There,
the polyclonal filter filters out beads that are estimated to have more than one template per ISP. Due to the nature of the protocol there always be some beads with more than one template on them. However, for a good quality run the "polyclonal" figure should be below 10%. Generally, high polyclonal figures isuggest ePCR titration issue.
See also "ISP Identification Summary" on the Ion Community:
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Old 12-21-2011, 05:12 AM   #3
Location: Guilford, CT and S.F., CA

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I concur, ionman. If you've upgraded to software v1.5.1 (and you should! lots of great improvements in there), here is an updated description of the ISP summary table, including the changes made to improve estimation of polyclonality:

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Old 12-23-2011, 12:08 PM   #4

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Most of the 1.5 reports seem to have higher polyclonal numbers than previous versions. I think they have changed the way it filters a couple times, so you can't compare to older reports.
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