Documents & Support

Why is my Ion AmpliSeq library concentration high? Product FAQ

Answer

Here are possible causes and solutions:
- The sample DNA or RNA may have been misquantified. We recommend requantifying the DNA or RNA. See the user guide for your library preparation method.
- More than 100 ng of sample DNA or RNA may have been used. We recommend decreasing the number of amplification cycles by adding less DNA or RNA.
- If you are using the Agilent 2100 Bioanalyzer instrument, the markers may have been misassigned. We recommend ensuring that the markers are correctly assigned.
- The library may have been contaminated by high molecular weight DNA. We recommend one of the following:
a) Removing less supernatant in the first-round (0.5X) purification and to ensure that the bead pellet is not disturbed.
b) Increasing the AMPur XP Reagent volume from 25 µL (0.5X) to 35 µL (0.7X) in the first-round purification.
- Inserts may have concatemerized during the ligation steps. We recommend reducing nucleic acid input, requantifying samples with a Qubit Fluorometer, or reducing the target amplification cycle number.

Answer Id:: E18935

Was this answer helpful?

Yes No

Thank you for your response

pEF6/V5-His TOPO TA Expression Kit Manual / Product Insert

  • Version: 28 June 2010
Catalog #

Brochure: Introducing the new iBright CL1500 and FL1500 Imaging Systems Product Literature

User Guide: Ion Torrent CarrierSeq ECS Kits Manual / Product Insert

  • Version: A.0
Catalog #

Rapid Isolation of Small Volume Plasmid DNA Using the 1-Liter General Purpose Centrifuge Product Literature

User Guide: Oncomine tumor specific panels Manual / Product Insert

  • Version: C.0

Isolating Genomic DNA from Whole Blood Product Literature

Handbook: Gibco Cell Culture Basics Handbook Product Literature

PichiaPink Expression System Manual / Product Insert

  • Version: MAN0000717 A.00 (22 Jan 14)

Product Reference Guide: Bioprocessing Portfolio Product Literature

Brochure: Western Blotting Laboratory supply checklist Product Literature

User Guide: Novex Pre-Cast gel electrophoresis guide Manual / Product Insert

  • Version: A.0
Catalog #

User Guide: Pichia Expression Kit Manual / Product Insert

  • Version: B.0
Catalog # K171001
Results per page
    spinner