The iCS-digital TM PSC 20q-only

The iCS-digital TM PSC 20q-only kit allows the detection by digital PCR of the most recurrent
genomic defect in human pluripotent stem cells (hPSCs).

The iCS-digital TM PSC 20q only kit
DNA extraction & quantification
1

DNA extraction & quantification

ddPCR reagent preparation
2

ddPCR reagent preparation

Droplet generation
3

Droplet generation

PCR
4

PCR

Droplet reader
5

Droplet reader

Analysis
6

Analysis

Species

Human

Cell types

hPSCs:
ESCs & iPSCs

Compatible instruments

QX100 and QX200
Droplet Digital PCR
Bio-Rad system

Storage

-20°C
upon reception

Size

20 tests

Coverage

The 20q11.21 gain is responsible for >20% of the recurrent abnormalities

Mosaicism

> 20%
(depending on sample quality)

The iCS-digital TM PSC 20q-only kit is based on the digital PCR (dPCR) technology and allows the reliable quantification of the sub-karyotypic 20q11.21 amplification. Gain of 20q11.21 copy-number variant (CNV) is detected in more than 20% of worldwide cultured human Pluripotent Stem Cells (hPSCs) 1-3 and represents 22.9% of the recurrent structural variants identified in hPSCs 4, making it the most common genomic abnormality in hPSCs.

Cells harbouring the 20q11.21 amplification display a selective advantage and can completely overtake the culture in few passages 5.

Detection of the 20q11.21 amplification using a dedicated technology is critical because most of these mutations fall below the size detection limit of conventional G-banding karyotyping (as illustrated on the right, panel B).

The kit also includes a validated normal genomic DNA control sample.

For Research Use Only

1, 2, 3, 4, 5 Refer to scientific publication number 1, 2, 3, 4 or 5 (bottom of page).

Sub-karyotypic 20q11.21 duplication in hPSCs (A) detected using the iCS-digital TM PSC 20q-only kit (sample 3) and (B) not identified by G-banding karyotyping method.
Images are for illustrative purposes only.

The iCS-digital TM 20q-only kit allows the fast and easy in-house analysis of the sub-karyotypic 20q11.21 amplification in hPSCs

The iCS-digital TM PSC 20q only
iCS-digital TM 20q-only kit content

Mix 20q that consists of :
- 1 specific probe assay
- 1 reference probe assay

Additional reagents required, but not included

ddPCR TM Supermix for Probes (No dUTP)
- Bio-Rad (#186023)

HindIII-HF Enzyme
- e.g. New England Biolabs (#R3104L)

1

Acquired genetic changes in human pluripotent stem cells: origins and consequences.

Halliwell J, Barbaric I, Andrews PW.
Nat Rev Mol Cell Biol. 2020 Dec;21(12):715-728.
2

Detecting Genetic Mosaicism in Cultures of Human Pluripotent Stem Cells.

Baker D, Hirst AJ, Gokhale PJ, Juarez MA, Williams S, Wheeler M, Bean K, Allison TF, Moore HD, Andrews PW, Barbaric I.
Stem Cell Reports. 2016 Nov 8;7(5):998-1012
3

BCL-XL mediates the strong selective advantage of a 20q11.21 amplification commonly found in human embryonic stem cell cultures.

Avery S, Hirst AJ, Baker D, Lim CY, Alagaratnam S, Skotheim RI, Lothe RA, Pera MF, Colman A, Robson P, Andrews PW, Knowles BB.
Stem Cell Reports. 2013 Oct 31;1(5):379-86
4

Recurrent Genetic Abnormalities in Human Pluripotent Stem Cells: Definition and Routine Detection in Culture Supernatant by Targeted Droplet Digital PCR.

Assou S, Girault N, Plinet M, et al.
Stem Cell Reports. 2020;14(1):1-8
5

Concise Review: Assessing the Genome Integrity of Human Induced Pluripotent Stem Cells: What Quality Control Metrics?

Assou S, Bouckenheimer J, and De Vos J.
Stem Cells. 2018; 36: 814-821

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