DNA sample preparation for epigenetic qPCR

Guideline for DNA Sample Collection
for Epiontis’ Epigenetic qPCR Immunomonitoring Service
DNA Isolation
To isolate genomic DNA from cells, tissue or whole blood the "DNeasy Blood & Tissue Kit"
from Qiagen (Cat. No. 69504) is recommended. A pdf version of the „DNeasy Blood & Tissue
Handbook“ is available online at www1.qiagen.com. Please refer to the handbook for
purification protocols and sample starting amounts.
For a high yield DNA isolation, please start with about 100 µl blood or a similar amount of
cells as suggested in the protocol, but replace the 100 µl elution buffer with 50 µl of elution
buffer, and use this amount to re-elute the column 3 times.
We recommend in general sending an amount of 10 µg DNA per sample in a volume of 50 µl
or less. This relatively high amount allows a re-measurement of your samples (at no
additional cost) in case a QC parameter for a plate run is not met or if a sample has an
unusually low target cell content.
If sample amounts are limited, please refer to the table on the second page of this guideline
for minimum DNA inputs that should be provided.
Shipping to Epiontis (Germany)
To avoid any customs delays, it is recommended to specify samples in the cover or
consignment letter as follows:
genomic DNA of human origin,
non-toxic, non-hazardous, non-infectious,
for research purpose only,
commercial value: 1 US $ / 1 Euro.
DNA samples can be sent at 4°C using cool packs, shipment on dry ice is not
Use clearly labelled safe lock tubes additionally sealed with parafilm to avoid leakage.
Please ship samples to:
Epiontis GmbH
Attention Julia Oppatt
Rudower Chaussee 29
12489 Berlin
Please notify Epiontis prior to shipment to check for German holidays and provide the
carrier’s tracking number as soon as the samples have been shipped (see email
below). Epiontis will monitor the progress of shipment and confirm sample receipt.
Version No.: 05
Replaces Version No.: 04
QMF 508-1e
Page: 1 of 2
Minimum DNA inputs
Cell Type
T cells
Regulatory T cells (Treg)
B cells
Th17 cells
T Helper cells
Cytotoxic T cells
Neutrophil Granulocytes
CCR-6 positive cells
Granulolysin positive cells
NK cells
TFH cells
Typical Level in Blood of
Healthy Donors [%]
DNA input amount
570 ng
2.6 µg
780 ng
3.8 µg
300 ng
480 ng
540 ng
2.5 µg
2.1 µg
2.6 µg
300 ng
620 ng
In more than 98% of measurements the minimum input will ensure measurements in the
validated assay range with standard precision if the sample contains at least half of the
typical target cell level observed with the epigenetic assay in blood of healthy donors.
Julia Oppatt
Manager Analytical Laboratory
[email protected]
Version No.: 05
Replaces Version No.: 04
Epiontis GmbH
Rudower Chaussee 29
12489 Berlin. Germany
QMF 508-1e
Fax: +49 30 6392 3476
Page: 2 of 2