DNA Extraction

Eurofins GeneScan Technologies GmbH

Engesser Str. 4
79108 Freiburg, Germany
Phone: + 49-(0)761-5038-100
Fax: + 49-(0)761-5038-111
DNAExtractor [email protected]
www.eurofins.com/kits

Fat
Kit for the extraction of DNA
from oil, fat and emulsifier (lecithin)

Cat. No. 5224700710

DNAExtractor Fat _ID2016 V8 17.11.2017 © 2017 Eurofins GeneScan Technologies GmbH, all rights reserved.

DNAExtractor Fat
Kit for the extraction of DNA
oil, fat and emulsifier

Cat. No. 5224700710

Table of Contents: DNAExtractor Fat Kit

1. INTRODUCTION 4 Kit for the extraction of DNA
from oil, fat and emulsifier
2. COMPONENTS OF THE KIT 5

3. REAGENTS/EQUIPMENT NOT INCLUDED 6

4. PREPARATION OF THE KIT’S REAGENTS 7

5. PROCEDURE 7
5.1 Equipment 7
5.2 DNA extraction 8

6. PRODUCT WARRANTIES AND SATISFACTION

GUARANTEE 13

7. IMPORTANT NOTES 15

8. TECHNICAL SERVICE 15

Kits, their components and instructions for use are

subject to alterations.

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 DNAExtractor Fat
Kit for the extraction of DNA
oil, fat and emulsifier

Cat. No. 5224700710

1. Introduction 2. Components of the Kit

This extraction kit is designed for the extraction of DNA The DNAExtractor Fat kit contains reagents for
for subsequent PCR testing. It yields DNA of high quality approx. 50 DNA extractions.
and purity.

The test kit is suitable for the extraction of DNA from oil, Store at room temperature
fat and emulsifier, e.g. lecithin samples. 1. (2x) Buffer EFG 50 mL

Store at – 4°C
2. (2x) Glycogen solution
(20 µg/µL) 2 x 100 µL (200µL)
The kit does not contain the solvents (ethanol,
isopropanol) necessary for extraction for freight 3. (1x) 100x TE stock buffer 20 µL
security reasons. (1000 mM Tris-Cl, 100 mM EDTA, pH 8.0)

For DNA Cleaning Columns see cat. nos

5224700310 (100 DNA Cleaning Columns),
5224700315 (5 x 100), 5224700311(10 x 100)

Important Note: Never store solutions and materials for

DNA extraction together with samples or amplicons.
Never use them in areas where PCR results are
interpreted (i.e. where gel electrophoresis & photo
documentation are performed).

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DNAExtractor Fat
Kit for the extraction of DNA
oil, fat and emulsifier

Cat. No. 5224700710

3. Reagents/Equipment not included 4. Preparation of the kit’s reagents

− Unpowdered (!) gloves Store the kit at –20°C until opened for the first time, if
− Ethanol p.a. (70 - 75% in H2O (sterile)) received in frozen condition. After first thawing, follow
− Chloroform p.a. the specific instructions regarding the correct storage
− Isopropanol p.a. (80 - 100% in H2O (sterile)) under chapter 2. Before use warm up the reagents for
− Water, DNase-free DNA extraction to room temperature. Dilute 100x TE
− 1 x TAE buffer stock buffer 1:1000 to 0.1x final concentration.
(40 mM Tris-acetate, 1 mM EDTA, pH 8.0) Avoid frequent thawing and freezing cycles, e.g. by
− n-Hexane p.A preparing aliquots at first use of the reagents kept
− Agarose frozen.
− Gel loading buffer
− EtBr (Ethidium bromide) 5. Procedure
− DNA Length Standard
− DNA Amount Standard 5.1 Equipment
− Tubes 50 mL
− Reaction tubes 1.5 mL and 2 mL
− Centrifuge (>4000 x g) suitable for 50 mL tubes
− Micro-centrifuge (>10.000 x g) for 1.5 and 2 mL tubes
− Blender / Gyrator / Vortex
− Heating oven / water bath (65°C)
− Thermocycler
− Gel electrophoresis equipment
− Transilluminator and polaroid camera or video system
− Micropipettes (variable 1-1000 µL)
− Filter tips

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 DNAExtractor Fat
Kit for the extraction of DNA
oil, fat and emulsifier

Cat. No. 5224700710

5.2 DNA extraction

The extraction of DNA from the sample is a crucial step, 5.2.1 Pretreatment of samples
and in order to guarantee optimal quantity and purity, 1. Warm up the lecithin samples in the microwave.
different methods are to be used depending on the Before starting the microwave: remember to
sample matrix. This kit is designed for fatty and oily loosen the cap of the bottle and remove any
samples and emulsifiers like lecithin. attached metal!!
2. Heat the vessels using medium heat (appr. 600 W or
Each sample should be analysed in duplicate.
lower) for 10 sec. If the lecithin is not liquid enough
It is recommended to perform an extraction control for
then repeat the procedure. Please note: You should
each set of samples extracted simultaneously, i.e. a be able to hold the vessel in your hand without
complete DNA extraction without sample material. getting hurt!! (Heat to max. 60°C)

This protocol was developed for the isolation of DNA 3. At the time when the sample is fluid, tighten the cap
from emulsifier/lecithin, oil and fat for PCR. again and gently agitate the lecithin by inverting the
All steps except otherwise indicated should be bottle preferentially with an overhead shaker (e.g. for
performed at room temperature. 20 min at 6 rpm) to homogenize the lecithin.

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DNAExtractor Fat
Kit for the extraction of DNA
oil, fat and emulsifier

Cat. No. 5224700710

5.2.2 Protocol for DNA extraction 8. For extraction controls, add 20 µL control DNA (e.g.
calf thymus or salmon sperm DNA) to the control
1. Weigh in 2.5 g of lecithin, fat or oil in a suitable tube. vessels.
(50 ml centrifuge tube). Preferentially extract all
9. Add 500 µL of chloroform and mix well: vortex or
samples in duplicate.
shake vigorously.
2. Prepare two tubes without sample material for
10. For the separation of organic and aqueous phases
extraction controls.
centrifuge for 10 min at >10.000 x g. Transfer the
3. Add 15 mL of hexane and 2 mL of Buffer EFG and mix aqueous upper phase into a new sterile 1.5 mL
vigorously under a ventilation fume hood until the reaction vessel.
sample material is dissolved, preferentially with an
11. Add 4 µL of glycogen solution and 0.8 volumes of
overhead shaker for about 15 min, alternatively vortex
80% - 100% isopropanol. Mix completely by inverting
for 10 – 20 sec.
the tube.
4. Centrifuge 10 - 15 min at 3 - 4000 x g until the phases
12. Incubate for 30 min at room temperature.
have separated. If no separation of phases was
achieved, add another 500 µL of Buffer EFG, mix 13. Precipitate the extracted DNA by centrifugation for
vigorously and centrifuge again. 10 min at 12.000 – 20.000 x g.
5. Transfer the aqueous phase (from the bottom of the 14. Carefully discard the supernatant (in most cases a
reaction tube; carry-over of intermediate phase is pellet of DNA should be visible at the bottom of the
tolerated) into a sterile 2 mL reaction vessel. reaction vessel).
6. Centrifuge the 2 mL vessel with the aqueous phase at 15. Add 500 µL 70% - 75% EtOH to the pellet and mix
room temperature for 10 minutes at 10.000 x g. by vortexing for at least 10 sec.
7. Transfer 1mL of the lower phase (excluding any 16. Precipitate the extracted DNA by centrifugation for
intermediate phase and pellet) in a new 2 mL vessel. 5 min at 12.000 – 20.000 x g.
17. Carefully discard the supernatant.

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 DNAExtractor Fat
Kit for the extraction of DNA
oil, fat and emulsifier

Cat. No. 5224700710

18. Centrifuge again and remove all remaining ethanol 6. Product Warranties and Satisfaction
with a pipette. Guarantee
19. Allow pellet to dry at room temperature for a
Eurofins GeneScan Technologies GmbH (“GeneScan”)
minimum of 5 minutes (if buffer crystals are visible,
repeat the washing step: addition of 70% - 75% warrants the products manufactured by it will be free of
ethanol followed by centrifugation; remove the defects in materials and workmanship when used in
supernatant carefully). accordance with the applicable instructions for a period
equal to or shorter of one year from the date of shipment
20. Dissolve the DNA pellet (in most cases visible) in of the product(s) or the expiration date marked on the
25 - 50 µL 0.1x TE buffer, either with buffer pre- product packaging under the storage conditions,
warmed to 65°C or incubation overnight at 4°C. recommended in the instructions and/or on the package.
21. If included/desired, apply the whole DNA extract to Application protocols published by GeneScan are
a DNA cleaning column (cat. no. 5224700310). intended to be only guidelines for the buyers of the
products. Each buyer is expected to validate the
applicability of each application protocol to his individual
application. GeneScan makes no other warranty,
expressed or implied. There is no warranty of
merchantability or fitness for a particular purpose.
GeneScan’s sole obligation with the respect to the
foregoing warranties shall be, at its option, to either
replace or to refund the purchase price of the product(s)
or part thereof that proves defective in materials or
workmanship within the warranty period, provided the
customer notifies GeneScan promptly of any such
defect. GeneScan shall not be liable for any direct,
indirect or consequential damages resulting from
economic loss or property damages sustained by buyer

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DNAExtractor Fat
Kit for the extraction of DNA
oil, fat and emulsifier

Cat. No. 5224700710

or any customer from the use of the product(s). A copy

of Eurofins GeneScan Technologies GmbH terms and 7. IMPORTANT NOTES
conditions can be obtained on request, and is also
provided with our product/price lists. Registered names, trademarks, etc. used in this
document, even when not specifically marked as such,
are not to be considered unprotected by law.
According to national regulations and standards, see
MSDS for details.

8. TECHNICAL SERVICE
If you have any questions or experience any difficulties
regarding this kit or Eurofins GeneScan Technologies
products in general, please do not hesitate to contact us.
Eurofins GeneScan Technologies customers are a major
source of information regarding advanced or specialized
uses of our products. This information is helpful to other
scientists as well as to the researchers at Eurofins
GeneScan Technologies. We therefore encourage you
to contact us if you have any suggestions about product
performance or new applications and techniques. For
technical assistance and more information please call
the Eurofins GeneScan Technologies Technical Service
Department or your local distributor.

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