Is the Ion Library Equalizer Kit compatible with Ion Ampliseq libraries?
The Ion Library Equalizer Kit (Cat. No. 4482298) is recommended for use with Ion AmpliSeq libraries and provides an alternative to library quantification methods by using bead-based technology to normalize the final library concentration to ~100 pM.
The Ion Library Equalizer Kit is fast and cost-effective compared to traditional quantification methods; however, it may be not be the right choice for all users. Briefly, the library is amplified with the Ion Equalizer Primers, captured onto Equalizer Beads, and the normalized library is eluted from the beads using a specially formulated Equalizer Elution Buffer. The final library is normalized to ~100 pM, but there is no quality control information (e.g., measured concentration or size distribution) that can be obtained, which is possible if using the recommended library quantification kits: Ion Library Quantitation Kit (qPCR), Qubit dsDNA HS Assay Kit (Qubit 2.0 Fluorometer), or Agilent High Sensitivity DNA Kit (Agilent Bioanalyzer instrument).
Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.
My custom design has low predicted coverage, what can I do to improve it?
If you've reviewed your design and are not satisfied with the results, please click on the Not happy with this design? Let us help link to have an Ion Ampliseq team member contact you about additional options for your design.
Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.
How do I get started with the Ion AmpliSeq Designer?
Please visit the Ion AmpliSeq Designer website (www.ampliseq.com) and log in with your Thermofisher username and password. Select the Help tab to review training videos and documentation, including pipeline details, troubleshooting guides, and frequently asked questions.
Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.
How should I dilute my library for template preparation? How much volume of the diluted library is required?
For template preparation, Ion AmpliSeq libraries are diluted to 100 pM, and the volume required for template preparation will vary depending on the template preparation kit used. Please see the Ion AmpliSeq Library Preparation User Guide for details regarding library dilutions and input into template preparation.
Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.
How long can I store my diluted Ion Ampliseq library?
Long-term storage of diluted libraries is not recommended and can result in decreased/suboptimal performance due to the adherence of DNA to the tube. In general, it is best to make fresh dilutions from the library stock for template preparation as needed. Libraries diluted for template preparation may be stored in a sealed plate or 0.2 mL PCR tube at 4-8 degrees C for up to 48 hours.
Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.