Product Description

PicoPLEX® technology- used to detect copy number variation in single cells on arrays is now available in a single tube format for use on Illumina NGS platforms.. PicoPLEX WGA has long been used by IVF clinics worldwide for pre-implantation genetic screening and diagnosis in detecting chromosomal aneuploidies and copy number variations.
PicoPLEX DNA-seq kit streamlines library preparation; the entire process is performed in a single tube or well – reducing error and contamination, speeding time to results, and reducing costs. PicoPLEX DNA-seq kit contains 48 reactions and includes everything necessary to convert 48 individual cells or DNA (6 pg to 60 pg) to NGS libraries, including dual barcodes. Barcoding oligonucleotides are provided in a single-use microwell plate.

Advantages

• Reduce ambiguity: highly reproducible CNV and aneuploidy detection
• Reduce workflow: from a single cell to a sequencing-ready library in three steps
• Reduce cost: a single kit contains everything needed to prepare a sequencing-ready library
• Reduce contamination and error: library prep in a single tube or well, no transfers necessary.
• Reduce time to results: Illumina NGS libraries prepared in less than 3 hours.

Specification

Store at –20 °C.
Guaranteed for 12 months at -20°C in a constant temperature freezer.

Kit selection guide

Performance

Highly Reproducible CNV Detection Over the Entire Genome

Amplified libraries from 11 individual flow-sorted H929 cells were sequenced on an Illumina MiSeq® and downsampled to 250,000 total reads. Thirty-five base single-end reads were mapped to human over the entire genome.

Highly Reproducible CNV Detection in Chromosome 1

In the each of the individual H929 cells (see previous slider), a 22 Mb loss and 30 Mb gain were consistently detected in chromosome 1 as indicated by the pink or blue bars.

PicoPLEX DNA-seq Kit Provides Aneuploidy Calls Concordant with Arrays

PicoPLEX DNA-seq Kit was tested for equivalency
to 24sure™ arrays (Illumina) by using the same
single embryo cell library for array and NGS testing. Libraries were pooled in groups with 48 compatible bar codes, and sequenced in a 2×100 MiSeq v2 PE experiment.
Sequencing results were concordant with array results and sequencing showed a terminal gain in 2p (shown in figure) and a terminal loss in 12q (data not shown), which were consistent with an undetected translocation in the paternal DNA confirmed by high resolution FISH analysis. (Data provided by GIVF).

Individual Library Quantification Eliminated

Flow-sorted unsynchronized H929 cells were amplified, pooled with constant volume, and loaded onto a MiSeq v3 flow cell. Quantification of the libraries was unnecessary before pooling due
to the highly reproducible amount of product
produced by PicoPLEX DNA-seq reactions. The
columns which provided no reads were wells in
which the cell was absent.

PicoPLEX DNA-seq Kit Library Preparation Workflow for Illumina NGS Platforms

PicoPLEX DNA-seq Kit Single-Tube Workflow

PicoPLEX DNA-seq Kit Technology

PicoPLEX DNA-seq Kit uses the same technology as the WGA kit: cells are lysed, quasi-random primers pre-amplify the DNA selectively, and a final PCR amplification adds the Illumina barcodes.

FAQ

Ordering Information

CAT. NO. R300381, 48 reactions.
The kit contains everything needed to do 48 reactions, including 48 dual indexes in a single use 96-well plate.

Download

PicoPLEX DNA-seq Instruction Manual

PicoPLEX DNA-seq Product Sheet

PicoPLEX DNA-seq Quick Protocol

Dual Index Plate Map

SDS PicoPLEX DNA-seq Kit