Equine oral samples collected and stabilized with Performagene™
are a reliable source of DNA for SNP genotyping on the Illumina®
Equine SNP70 Beadchip
C. Acres1, R. Iwasiow1, M. Tayeb1, W.A. van Haeringen2 and M. de Groot2
1 DNA Genotek Inc., Ottawa, Canada www.dnagenotek.com
2 VHLGenetics, Netherlands www.vhlgenetics.com

Introduction

DNA Conc.
(ng/μL)

A260/A280

Call rate

FarmAge

Gender

9.2

1.50.990

Farm A

16M

40.9

1.70.892

Farm A

12M

1.4

1.30.982

Farm A

15F

4.1

1.80.982

Farm A

18

F18.6

1.60.941

Farm A

6M

43.5

1.80.987

Farm A

12M

16.8

1.70.987

Farm A9M

15.9

1.70.964

Farm A

17F

17.7

1.70.985

Farm A

20F

24.2

1.70.969

Farm A

12F

13.4

1.70.984

Farm A

22F

18.5

1.60.929

Farm A

7M

7.4

1.60.978

Farm A

1.5M

37.6

1.40.967

Farm A4M

78.4

1.80.936

Farm B28M

6.7

1.30.987

Farm C

11M

15.2

1.60.990

Farm C

18M

22.5

1.60.989

Farm C

19F

Extraction and analysis was performed by VHLGenetics®. Of the samples collected,50% were not included in the subsequent analysis as they were dirty samples.

7.5

1.30.973

Farm C

14M

18.4

1.40.907

Farm C

16M

3.1

1.40.987

Farm C

10FExtraction

125.4

1.90.989

Farm D

6M

11.9

1.50.965

Farm D

5M

Oral samples collected and stabilized with Performagene™ has provento be an easy and cost-effective way to collect, transport and extract
DNA for downstream analysis.
In this study we investigate the performance of DNA extracted
from oral samples collected using Performagene on the Illumina®
Equine SNP70 Beadchip.

Methods and results

3451122Collection, stabilization and transport
Oral samples were collected and stabilized using Performagene from 48 horses of varying
breeds across 4 different farms in Ontario, Canada. Horses of both genders aged 1.5 years
to 28 years were sampled.

All samples were vortexed thoroughly to obtain an optimal suspension of the sample.
The sbeadex™ kit from LGC was used to extract DNA from the samples: 100 μL of oral sample
was transferred to a 96-wells plate. 100 μL of binding buffer, containing magnetic beads
was subsequently added to the samples together with 200 μL lysis buffer with proteinase K.
Samples were washed twice and the DNA was finally dispensed in the elution buffer provided
by manufacturer.

Genotyping arrays

Quality control

Highlights

DNA concentration was measured by PicoGreen®. The range of concentration was 1.4 ng/μL
to 125.4 ng/μL.
A260/A280 was performed. The range was between 1.3 and 1.9.
The purified DNA quality was assessed using an agarose gel. All samples were of good quality.
This is a representation of the quality.

23 kb

12Superior samples
Proven performance

345678

Table 1: Summary of analyzed samples. (> 0.89)

2 μL of DNA was used on each chip and standard protocols from Illumina were followed
to yield the obtained results. Data for samples with a call rate > 0.89 is reported.

Performagene is a non-invasive, all-in-one system to collect, stabilize and transportoral DNA samples from horses.
Using VHLGenetics extraction platform, high quantity and quality DNA
was extracted from Performagene oral samples.
Genotyping results demonstrate that oral samples
can be a good source of DNA for successful
genotyping on the Illumina Equine SNP70 Beadchip.
Data suggests that a robust extraction may berequired to remove inhibitors that were present
in a subset of samples and which in most cases
could be identified prior to processing as“dirty” samples. Other extraction methods
are being evaluated to clean these samples.
Performagene samples processed byVHLGenetics provide cost-effective end-to-end
solutions for DNA sample management fromhorses, including simplifying collection,transportation, extraction and analysis.

Performagene is For Animal Use Only.
Some DNA Genotek products may not be available in all geographic regions.
Performagene is a trademark of DNA Genotek Inc.
All brands and names contained herein are the property of their respective owners.
© 2016 DNA Genotek Inc., a subsidiary of OraSure Technologies, Inc., all rights reserved.
Patent (www.dnagenotek.com/legalnotices) MK-00560 Issue 1/2016-01

www.dnagenotek.com • support@dnagenotek.com