Genetic and genomic analysis from tissue samples requires the extraction of high quality DNA. Mechanical disruption methods such as bead milling provide high yield from tissue samples, but cause damage to the nucleic acids. Purely enzymatic methods such as proteinase K digestion can extract nucleic acid without damage, but require long incubation times, often proceeding overnight, and without approaching the yields achieved by mechanical disruption techniques. Thus a method is needed which can provide a rapid extraction of high yield, high quality DNA from tissue samples. See the new method.

1. Dominic O’Neil, Stefanie Schröer, Tanya Sperling, Markus Sprenger-Haussels
QIAGEN Strasse 1, 40724 Hilden, Germany
11/2017
Rapid Purification of High-Yield, High-Quality DNA from
Tissue Samples Using the QIAamp®
Fast DNA Tissue Kit
Sample to Insight
Introduction
Genetic and genomic analyses from tissue samples require high-quality DNA. Mechanical disruption methods, such as
bead milling provide a high yield from tissue samples, but cause damage to nucleic acids. Purely enzymatic methods
such as proteinase K digestion can extract nucleic acid without damage, but require long incubation times, and without
approaching the yields achieved by mechanical disruption techniques. Therefore, a method is needed which can provide
rapid purification of high-yield, high-quality DNA from tissue samples.
Method
The QIAamp Fast DNA Tissue Kit combines mechanical, chemical and enzymatic disruption of tissue samples to provide rapid,
high-yield DNA purification – without significant damage to the nucleic acid. Tissue is mechanically disrupted by vortexing with a
specially shaped milling bead. The lower power of a vortexer compared to a bead mill prevents DNA shearing. Low chaotropic
conditions support mild sample lysis and provides synergy for the simultaneous proteinase K digestion and mechanical shearing.
Results
DNA purified from fresh-frozen and stabilized tissue samples using the QIAamp Fast DNA Tissue Kit showed up to a 10-fold
increase in yield compared with standard proteinase K digestion or mechanical milling alone. Disruption was complete in
15 minutes for soft tissues (e.g., liver, kidney), and within 30 minutes for tougher tissue (e.g., stabilized lung, trachea samples).
DNA was also efficiently purified from fatty tissues (e.g., brain) and fibrous tissues (e.g., muscle).
Summary
The QIAamp Fast DNA Tissue Kit enables superior DNA purification from tissue compared with existing methods. Technologies
including next-generation sequencing make the purification and analysis of DNA from tissue samples increasingly important.
Therefore, a method that provides convenient and efficient purification of DNA supports the routine genomic analysis
facilitated by these technologies.
Tissue samples are added to tubes containing specially shaped beads that homogenize
tissue during mechanical agitation. Mechanical lysis proceeds in a gentle chaotropic
solution, in which the components for enzymatic digestion are already present. The
chaotropic buffers denature proteins to make them more accessible to proteinase K.
Synergistically, proteinase K digestion softens tissue, making it more susceptible to
mechanical disruption. The mechanical disruption in turn liberates additional material
for the chaotropic salts and proteinase K. The three components together provide rapid
tissue homogenization.
The method was developed for various downstream applications, including (q)PCR and
next-generation sequencing.
Combined Enzymatic, Chemical and Mechanical Tissue Lysis
Combination Lysis Enables Rapid, High-Yield DNA Purification
Rat and mouse tissues – including fatty tissue (e.g., brain), fibrous tissue (e.g., heart) and tough tissue (e.g., trachea) were
processed using the QIAamp Fast DNA Tissue Kit in comparison to kits using enzymatic homogenization alone.
Yields (determined by dsDNA-specific assays) of all tissues were up to 10-fold higher using the QIAamp Fast DNA Tissue Kit.
The QIAamp Fast DNA Tissue Kit can be used with vortexers, the TissueLyser LT or TissueLyser II with comparable performance.
Up to 10-fold higher yields with the QIAamp Fast DNA Tissue Kit, compared with enzymatic lysis alone.
DNA was purified from 15 mg of stabilized rat kidney, lung and liver and 1 cm fresh-frozen mouse tail tip. The QIAamp
Fast DNA Tissue Kit was compared with spin-column-based purification kits from other suppliers (with solely mechanical or
enzymatic lysis or a less optimized combination of mechanical and enzymatic homogenization). In all cases, the QIAamp
Fast DNA Tissue Kit gave the best combination of time and yield.
Efficient Purification from Tough, Fibrous or Fatty Tissues
Conclusions
The rapid and gentle lysis provided by the the QIAamp Fast DNA Tissue Kit demonstrated superior results compared with
existing methods.
• DNA yield was up to 10-fold higher with the the QIAamp Fast DNA Tissue Kit than with other enzymatic and/or mechanical
homogenization techniques.
• The QIAamp Fast DNA Tissue Kit has been shown to work with a wide variety of tissue types – including fatty, fibrous
and tough tissues.
• DNA integrity (in terms of molecular weight) extracted from tissue is improved with the QIAamp Fast DNA Tissue Kit,
compared with methods from other suppliers.
• Since DNA purification is a major step in almost all genetic and genomic applications, the QIAamp Fast DNA Tissue
Kit offers a new approach for tissue investigation.
The QIAamp Fast DNA Tissue Kit is intended for molecular biology applications. This product is not intended for the
diagnosis, prevention, or treatment of a disease.
For up-to-date licensing information and product-specific disclaimers, see the respective QIAGEN kit handbook or user
manual. QIAGEN kit handbooks and user manuals are available at www.qiagen.com or can be requested from QIAGEN
Technical Services or your local distributor.
Trademarks: QIAGEN®
, Sample to Insight®
, QIAamp®
(QIAGEN Group). Registered names, trademarks, etc. used in this document, even when not specifically
marked as such, are not to be considered unprotected by law.
© 2017 QIAGEN, all rights reserved. PROM-11655-001
DNA Quality is Improved With Combination Lysis
DNA was purified using either the QIAamp Fast DNA Tissue Kit or methods from other suppliers. DNA quality was
analyzed using pulsed-field gel electrophoresis (PFGE). DNA purified using the QIAamp Fast DNA Tissue Kit showed
highest integrity. The speed of purification preserves DNA integrity compared with enzymatic digestion alone, while the
comparatively gentle mechanical agitation improves DNA integrity over other mechanical homogenization methods.
High-quality DNA with the QIAamp Fast DNA Tissue Kit. Genomic DNA was purified from 15 mg stabilized rat kidney and lung and 1 cm fresh-frozen
mouse tail tip using either the QIAamp Fast DNA Tissue Kit (with vortex, TissueLyser II or TissueLyser LT) or a spin-column-based kit (with solely mechanical
or enzymatic or a combination of mechanical and enzymatic homogenization). Equal amounts of DNA were analyzed using PFGE, except for the mouse
tail sample from Kit C, where due to low yields, the maximum possible amount of eluate was analyzed. E: Enzymatic lysis; M: Mechanical lysis;
M+E: Mechanical and enzymatic lysis; TL: TissueLyser II or LT; V: Vortexer.
Ballcone beads for gentle tissue disruption. QIAamp Fast DNA Tissue procedure.
Consistently high yields of DNA and rapid purification times with the QIAamp Fast DNA Tissue Kit.
QIAamp Fast DNA Tissue Procedure
Disrupt and lyse
Sample
Bind
Wash
Elute
DNA
Kidney
Lung
Enzymatic lysis only
QIAamp Fast – vortexer
Mechanical lysis only
QIAamp Fast – TissueLyser LT
Combined enzymatic/mechanical
QIAamp Fast – TissueLyser II
0
5
10
15
20
25
30
35
40
45
Yield (µg dsDNA)
Liver
Yield (µg dsDNA)
050100150200250300
Time (minutes)
050100150200250300
Time (minutes)
Improved performance
0
10
20
30
40
50
60
70
050100150200250300
Yield (µg dsDNA)
Mouse tail
Yield (µg dsDNA)
Time (minutes) Time (minutes)
0
5
10
15
20
25
30
35
0
5
10
15
20
25
050010001500
Ratheart
Ratspleen
Ratbrain
Ratesophagus
Rattrachea
Ratear
Ratfat
Rattail
Pig
ear
0.1
1
10
100
Yield (µg dsDNA)
QIAamp Fast – vortexer
QIAamp Fast – TissueLyser II
QIAamp Fast – TissueLyser LT
Enzymatic homogenization
145,5
97
48,5
23,1
9,42
6,55
4,36
194
Rat lung
145.5
97
194
LkbV TLM
145.5
97
48.5
23.1
9.42
6.55
4.36
194
Rat kidney
kb L
Kit A
QIAamp Fast
(M+E)
E
Kit B
M+E
Kit C
V TLM
Kit A
QIAamp Fast
(M+E)
E
Kit B
M+E
Kit C
V TL
QIAamp Fast
(M+E)
M
Kit
A
E
Kit
B
M+E
Kit
C
Mouse tail
145.5
97
48.5
23.1
9.42
6.55
4.36
194
Lkb