VeriSeq NIPT Method

The VeriSeq NIPT Method (Method) directs the automated processing of samples on the ML STAR. The method accomplishes the following processing steps:

•

Plasma Isolation—Transfers 1 ml of isolated plasma from a blood collection tube. The process logic creates a batch with the VeriSeq NIPT Assay Software. Each batch contains sample data, including the sample barcode, sample type, screen type, well position, and sex-reporting flag.

•

Cell-Free DNA (cfDNA) Extraction—Purifies cfDNA from 900 μl of plasma.

•

Library Preparation—Creates libraries from purified cfDNA that are ready for sequencing. The libraries include unique indexes for each sample in the batch.

•

Library Quantification—Determines cfDNA concentration using an intercalating fluorescent dye in a 384-well microplate format. The plate includes a labeled DNA standard curve and duplicates of each sample in the batch. The system uses the raw fluorescence readings from the microplate reader and calculates the sample concentrations based on the standard curve.

•

Pooling and Normalization—Combines libraries into single pools for sequencing. The system uses the previously determined concentrations to calculate the appropriate transfer volumes for each sample in the sequencer-ready pool.