The Ion AmpliSeq™ Transcriptome Human Gene Expression Kit enables the simultaneous measurement of the expression levels of over 20,000 human RefSeq genes in a single assay. Providing the ability to work directly with low input levels of total RNA derived from challenging samples such as formalin-fixed paraffin-embedded (FFPE) tissue or other samples with degraded RNA, the Ion AmpliSeq™ Transcriptome Human Gene Expression Kit is ideal for those interested in obtaining comprehensive, highly sensitive, and easily obtained gene expression measurements from a wide range of RNA sources.

The Ion AmpliSeq™ Transcriptome Human Gene Expression Kit offers:
• Flexible, straightforward library preparation – start with as little as 10 ng of total RNA from FFPE samples, cells, or other sources
• Representation of over 20,000 human RefSeq genes
• Wider dynamic range and improved detection sensitivity compared to conventional microarray analysis
• A complementary software plug-in for automated sequencing analysis that provides gene quantitation and reporting as well as quality control metrics
• Compatibility with Ion Xpress™ Barcode adapters allowing for multiplexed sequencing

Wider dynamic range for enhanced performance
With a wider dynamic range and improved gene detection compared to conventional microarray analysis, the Ion AmpliSeq Transcriptome™ Human Gene Expression Kit provides a clear solution to researchers needing enhanced performance for their gene expression needs. A simple workflow allows for sample-to-gene quantitation in less than 2 days, helping to reduce workload and increase throughput.

Straightforward library preparation and multiplex sequencing
Starting with as little as 10 ng of total RNA, libraries can be prepared in approximately 7 hours with only one hour of hands-on time. Total RNA is first processed with the included SuperScript™ VILO™ cDNA Synthesis kit, and the resulting cDNA is amplified using Ion AmpliSeq™ technology to accurately maintain expression levels of all genes.

The resulting libraries are ready for downstream template preparation using the Ion OneTouch™ 2 System or Ion Chef™ System and sequencing on the Ion Proton™ sequencing system. Libraries can be barcoded for multiplexing, allowing for flexibility, helping to reduce cost, and increase throughput, with the recommended 16 libraries per Ion 550 Chip and 8 libraries per Ion 540 or Ion PI chip. Other multiplexing options are also possible, depending on the experimental design and desired level of sensitivity.

Automatic data analysis with Torrent Suite™ plug-in
Using the complementary analysis plug-in available with Torrent Suite™ software, gene quantitation information is automatically compiled, minimizing the requirement for costly computational resources and bioinformatics expertise to process raw sequencing data into meaningful information. An easy to interpret, highly portable table is produced containing all genes interrogated and their respective expression levels, output that may be used with a variety of existing software applications for downstream analysis of microarray or next-generation sequencing data.

Note: Libraries made from this protocol yield fragments suitable for 200-base read sequencing.