Frequently Asked Questions

Are there any other kit size options?

Currently, we only have one kit size option, the 960 sample version. This kit is set up as 10 x 96 sample reactions, with a one-year shelf life, so all 960 do not need to be done at once. Note that it is still cost-competitive to sequence less than 96 samples at a time if you do not have enough samples to fill a plate.

Is it feasible to prepare less than 96 samples at once?

Yes. The kit is intended to process 10 batches of 96 samples, however, you could process less than the full 96 for each batch. It would still be cost-effective down to 8 samples per batch when compared to most other technology. You could also run samples as replicates, which allows for greater confidence in variant calling.

Are you be able to provide any kind of tech support?

Yes. Dr Azeem Siddique, iGenomX Director of RnD, is leading our technical support group and either he or one of his team will be available to help with any issues that may arise. For technical support, please contact [email protected] or let us know how we can help via the contact form.

What is the amount of the DNA needed?

50 ng input is recommended but we do have a protocol to go down to 1 ng.  If you would like the low input protocol, please let us know via the contact form

Is there a maximum input amount of DNA?

Adding more DNA typically improves the performance of all preps.  With that said, we have not tested above 100ng with the RIPTIDE technology.  We want to make sure there is enough flexibility for users to make if you want to increase the input, please just make sure you are adding an equimolar amount to each well in the plate.

How important is the input DNA quality and quantity? How important is the input DNA quality and quantity?How important is the input DNA quality and quantity?

Aim for inputs with 1kb or greater fragments if you can. Synthesized DNA (plasmids, gene constructs), PCR amplicons (long-range) etc. have all shown good results. Ultra-long molecules are not a problem at all. We can work with you on tweaking for more degraded samples.

Do you have any data sets or papers that can be shared, demonstrating the performance of your technology?

Yes, please see our Resources section on our website for scientific posters and data sets.

What is the shelf life of this kit?

One year

Is the RipTide;trade; kit compatible with any NGS platform?

The current version of the RIPTIDE™ kit is compatible with any Illumina sequencer and read length and can be tailored to other NGS platforms. Please contact us via the contact form for more information.

Can we reduce volume ratios for the reagents?

I would caution that there is one limiting component.  That is the streptavidin capture beads that capture the biotinylated products from the initial barcoding reaction.  Simply put, there are only so many spots on a bead to bind a molecule.  If you reduce the beads, you, therefore, reduce the number of molecules that are captured from a given sample.  Reducing molecule inputs into a PCR reaction could reduce library complexity and increase PCR duplication.  

Can you provide higher than 960 indexes?

Yes, we can and would be happy to work with you on this. Please let us know via the contact form on our website. Please click HERE to contact us for more information.

Are there any specific considerations for using this technology on a novaseq?

There should be no problem with the NovoSeq instrument as we are tailored to work on all Illumina Sequencers. You will, however, want to be aware of your level of barcode hopping inherent in the NovoSeq instrument.

Do you provide a free demultiplexing tool?

Yes, free open source software is provided for sample demultiplexing. Links to the software are can also be found in the resources section of our website under the "Software" subsection, as well as in our manual. Our tool lets you implement into any pipeline.

Can the RipTide kit take RNA as an input?

Yes, via a cDNA intermediary step.

What does the kit contain and which additional consumables are required for library prep (besides those necessary to generate input DNA).

All you need in addition to the kit are pipette tips, a magnetic stand and EDTA.  So everything else to go from DNA to size selected libraries are included in the kit.  

Why is it helpful to avoid fragmentation?

Fragmentation causes a large amount of information to be lost by destroying many of the very molecules that one tries to preserve. This is especially alarming for rare and small genes of interest, which will go undetected if fragmented

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