PicoPLEX™ WGA Kit

Whole Genome Amplification for Library Construction of Single Cells

picoplex

“SurePlex™ [PicoPLEX]  represents a major step forward in amplification technology…we selected SurePLEX due to its rapid protocol, highly representative amplification,and low allele dropout when compared to other amplification methods.”

–Nick Haan, founder and CEO of BlueGnome, an Illumina Company

Overview

The ability to amplify DNA to yield a highly reproducible library from a single cell is now possible! The PicoPLEX™ WGA kit  with patented technology* is designed and optimized for amplification of single copy genomic DNA  starting with a single cell.  The easy-to-use single tube protocol reduces handling errors, dramatically improves time to results and reduces background. PicoPLEX WGA Kit can be also be used with isolated gDNA amounts ranging  from less than 15 pg to 50 ng. Accepted for use in array CGH analysis as well as q-PCR.

*Protected by US Patent 8,206,913 and EU pending applications

PicoPLEX™ WGA Kits are used and trusted by leading providers in the IVF community such as BlueGnome for pre-implantation genetic screening and diagnostics (PGS/PGD).

  • Reproducible results from single cells (or <15 pg DNA)
  • Superior reproducibility of allele representation
  • Easy to use and automate
  • Unambiguous results at all resolutions, microarray and PCR

Workflow

picoplex_wga_tube_rev2

Learn more about PicoPLEX technology.

Amplification of Single Cells is Reproducible

Amplification of Single Cells is Reproducible
Amplification of Single Cells is Reproducible

Flow-sorted cancer cells were amplified with the PicoPLEX WGA Kit. Each of the cells (blue lines) amplified at the same rate, and resulted in a similar, predictable yield. The sample containing 0 cells (green lines; no template control) shows very low background.

Reproducible CNV Data

Reproducible CNV Data
Reproducible CNV Data

Single-blastomere biopsies were amplified using PicoPLEX WGA kit, labeled and hybridized to BlueGnome’s 24Sure arrays at Genesis Genetics Institute. Note clear indication of CNVs. In 2011, ESHRE clinical trials confirmed accuracy of karyotyping using PicoPLEX technology.

PicoPlex 3

PicoPlex 3
PicoPlex 3

Locus-specific q-PCR was used to quantify 48 loci in independent single-cell libraries. Data shown compares results of DNA from two individual samples amplified with (left to right) PicoPLEX WGA Kit , GenomePLEX® and GenomiPhi™ using 10 pg of DNA. More than 90% of the product from PicoPLEX produce identifiable human sequences and this was highly reproduced,

Store at -20 °C.

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

Recommended DNA inputs:

  • 1 – 10 human cells (e.g. blastomeres, polar bodies, trophoblastic cells, amniocytes, CTCs, cultured cells
  • 1,000 – 10,000 bacterial cells
  • Intact or fragmented, single- or double-stranded DNA

Maximum Sample Volume, 5 µL

Top Ten FAQs  

1. What applications are recommended for PicoPLEX WGA?
This kit is used for:

  • Copy number variation (CNV) analysis: superior for oligonucleotide and BAC aCGH
  • SNP genotyping
  • Mutation detection by PCR

2. Is there a non-stick wash buffer that can be used for single cell wash prior to using PicoPLEX WGA kit?
This has not been validated at Rubicon, but some customers have successfully used PVA (Polyvinyl Alcohol; e.g. 0.1%) in the PBS to wash/collect their cells. Additionally, your washing/collecting buffer (PBS) must be also Ca- and Mg-free.

3. Are there special requirements for flow sorting?
Yes, we strongly recommend not fixing, and using light scattering or phase contrast to sort or collect samples. Microscopic/visual confirmation of successfully sorted cells can be used to optimize sorting conditions.

4. What is the sample input volume for PicoPLEX WGA?
This kit will accommodate a 5 μl sample input volume per reaction (with a maximum of 2.5 μl of cell & buffer carryover).

5. Can the product of PicoPLEX WGA be taken straight to NGS?
No.

6. Sigma-Aldrich sells a GenomePlex WGA kit, is it the same technology as PicoPLEX WGA?
No.

8. What is the expected genomic locus representation and Loss of Heterozygosity (LOH) for PicoPLEX WGA products?
For reference purposes, see the data obtained from a third party in the table below.

SNP Genotyping Method Single-Cell  Success Rate NP Call Rates LOH
PCR 95% > 95% < 10%
Illumina Infinium SNP array 95% 50% – 60% 7% – 12%

9. Is the product of this technology single- or double-stranded?
The end product of PicoPLEX WGA is a mixture of single- and double-stranded DNA.

10. How rapid is PicoPLEX WGA?
PicoPLEX WGA lyses cells and amplifies the DNA to an end-point of 2 – 5 μg in < 3 hours.

PicoPLEX™WGA Kit is available in a 50 reaction size.

Order direct through our STORE

Order from New England BioLabs.

Order from one of our Distributors.

For larger volumes or custom applications, contact busdev@rubicongenomics.com

Performance

Amplification of Single Cells is Reproducible

Amplification of Single Cells is Reproducible
Amplification of Single Cells is Reproducible

Flow-sorted cancer cells were amplified with the PicoPLEX WGA Kit. Each of the cells (blue lines) amplified at the same rate, and resulted in a similar, predictable yield. The sample containing 0 cells (green lines; no template control) shows very low background.

Reproducible CNV Data

Reproducible CNV Data
Reproducible CNV Data

Single-blastomere biopsies were amplified using PicoPLEX WGA kit, labeled and hybridized to BlueGnome’s 24Sure arrays at Genesis Genetics Institute. Note clear indication of CNVs. In 2011, ESHRE clinical trials confirmed accuracy of karyotyping using PicoPLEX technology.

PicoPlex 3

PicoPlex 3
PicoPlex 3

Locus-specific q-PCR was used to quantify 48 loci in independent single-cell libraries. Data shown compares results of DNA from two individual samples amplified with (left to right) PicoPLEX WGA Kit , GenomePLEX® and GenomiPhi™ using 10 pg of DNA. More than 90% of the product from PicoPLEX produce identifiable human sequences and this was highly reproduced,

Specifications

Store at -20 °C.

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

Recommended DNA inputs:

  • 1 – 10 human cells (e.g. blastomeres, polar bodies, trophoblastic cells, amniocytes, CTCs, cultured cells
  • 1,000 – 10,000 bacterial cells
  • Intact or fragmented, single- or double-stranded DNA

Maximum Sample Volume, 5 µL

Resources

FAQs

Top Ten FAQs  

1. What applications are recommended for PicoPLEX WGA?
This kit is used for:

  • Copy number variation (CNV) analysis: superior for oligonucleotide and BAC aCGH
  • SNP genotyping
  • Mutation detection by PCR

2. Is there a non-stick wash buffer that can be used for single cell wash prior to using PicoPLEX WGA kit?
This has not been validated at Rubicon, but some customers have successfully used PVA (Polyvinyl Alcohol; e.g. 0.1%) in the PBS to wash/collect their cells. Additionally, your washing/collecting buffer (PBS) must be also Ca- and Mg-free.

3. Are there special requirements for flow sorting?
Yes, we strongly recommend not fixing, and using light scattering or phase contrast to sort or collect samples. Microscopic/visual confirmation of successfully sorted cells can be used to optimize sorting conditions.

4. What is the sample input volume for PicoPLEX WGA?
This kit will accommodate a 5 μl sample input volume per reaction (with a maximum of 2.5 μl of cell & buffer carryover).

5. Can the product of PicoPLEX WGA be taken straight to NGS?
No.

6. Sigma-Aldrich sells a GenomePlex WGA kit, is it the same technology as PicoPLEX WGA?
No.

8. What is the expected genomic locus representation and Loss of Heterozygosity (LOH) for PicoPLEX WGA products?
For reference purposes, see the data obtained from a third party in the table below.

SNP Genotyping Method Single-Cell  Success Rate NP Call Rates LOH
PCR 95% > 95% < 10%
Illumina Infinium SNP array 95% 50% – 60% 7% – 12%

9. Is the product of this technology single- or double-stranded?
The end product of PicoPLEX WGA is a mixture of single- and double-stranded DNA.

10. How rapid is PicoPLEX WGA?
PicoPLEX WGA lyses cells and amplifies the DNA to an end-point of 2 – 5 μg in < 3 hours.

Ordering Information

PicoPLEX™WGA Kit is available in a 50 reaction size.

Order direct through our STORE

Order from New England BioLabs.

Order from one of our Distributors.

For larger volumes or custom applications, contact busdev@rubicongenomics.com