MinElute and QIAquick DNA Cleanup Systems

The MinElute and QIAquick DNA Cleanup Systems are designed for fast cleanup of DNA fragments from PCR, other enzymatic reactions, and agarose gels. Using silica-gel–membrane technology, both MinElute and QIAquick kits deliver high-purity DNA suitable for use in all applications. MinElute kits deliver highly concentrated DNA using minimal elution volumes.

PCR Cleanup
	Size range	No. of samples in parallel	Format	Processing
MinElute PCR Purification Kit	70 bp – 4 kb	1–24	Spin columns	Spin or vacuum
MinElute 96 UF PCR Purification Kit	100 bp – 4 kb	96	96-well plates	BioRobot systems (automated) or QIAvac Multiwell (manual)
QIAquick PCR Purification Kit	100 bp – 10 kb	1–24	Spin columns	Spin or vacuum
QIAquick 8 PCR Purification Kit	100 bp – 10 kb	8–48	8-well strips	QIAvac 6S
QIAquick 96 PCR Purification Kit	100 bp – 10 kb	96	96-well plates	QIAvac 96
QIAquick 96 PCR BioRobot Kit	100 bp – 10 kb	96	96-well plates	BioRobot Systems

Gel Extraction
	Size range	No. of samples in parallel	Format	Processing
MinElute Gel Extraction Kit	70 bp – 4 kb	1–24	Spin columns	Spin or vacuum
QIAquick Gel Extraction Kit	70 bp – 10 kb*	1–24	Spin columns	Spin or vacuum

* DNA fragments smaller than 70 bp or larger than 10 kb should be extracted using the QIAEXII Gel Extraction System.

Cleanup of DNA from Enzymatic Reactions
	Size range	No. of samples in parallel	Format	Processing
MinElute Reaction Cleanup Kit	70 bp – 4 kb	1–24	Spin columns	Spin or vacuum
QIAquick DNA Cleanup System	Varies depending on kit	1–24	Spin columns	Spin or vacuum

Nucleotide Removal
	Size range	No. of samples in parallel	Format	Processing
QIAquick Nucleotide Removal Kit	Oligos: 17–40mers dsDNA: 40 bp – 10 kb	1–24	Spin columns	Spin or vacuum

Guide to Enzymatic Reaction Cleanup Using the QIAquick DNA Cleanup System
	QIAquick PCR Purification Kit	QIAquick Nucleotide Removal Kit	QIAquick Gel Extraction Kit
For DNA cleanup from the following reactions:
Alkaline phosphatase	YES	YES	YES
cDNA synthesis	YES	no	no
DNase, nuclease digestion	YES	YES	YES
Kinase:
DNA fragments	YES	YES	YES
Oligonucleotides	no	YES	no
Ligation	YES	YES	YES
Nick translation	YES	YES	YES
PCR	YES	no	no
Random priming	YES	YES	YES
Restriction digestion	YES	YES	YES
Tailing:
DNA fragments	YES	YES	YES
Oligonucleotides	no	YES	no
Specifications
Recovery:
Oligonucleotides	–	17–40mers	–
dsDNA	100 bp – 10 kb	40 bp – 10 kb	70 bp – 10 kb
Removal:
<10mers	YES	YES	YES
17–40mers	YES	no	no

Features and benefits

MinElute offers:

Very small elution volumes — highly concentrated DNA eluted with just 10 µl (see figure "Higher DNA Concentrations Obtained Using MinElute Kits")
Fast procedure — purification of PCR products and DNA from enzymatic reactions in 6 minutes, gel extraction in 18 minutes
Uniquely designed silica-gel–membrane spin technology
High recoveries — 80% of 70 bp to 4 kb fragments

Higher DNA Concentrations Obtained Using MinElute Kits

A 500 bp and a 1000 bp fragment purified using the MinElute Gel Extraction Kit and 3 different silica-based DNA purification kits from the suppliers indicated. Two microliters of each eluate was loaded onto a 1.5% agarose gel. M: markers.

QIAquick offers:

Up to 95% recovery of ready-to-use DNA
Fast procedure — purification of PCR products and DNA from enzymatic reactions in 5 minutes, gel extraction in 15 minutes
A system designed for different throughput needs
Cleanup of DNA up to 10 kb in 3 simple steps: just bind, wash, and elute

Principle

MinElute and QIAquick Kits contain a silica-gel– membrane assembly for binding of DNA in high-salt buffer and elution with low-salt buffer or water. The purification procedure removes primers, nucleotides, enzymes, mineral oil, salts, agarose, ethidium bromide, and other impurities from DNA samples. Silica-membrane technology eliminates the problems and inconvenience associated with loose resins and slurries. Specialized binding buffers are optimized for specific applications and promote selective adsorption of DNA molecules within particular size ranges.

MinElute 96 UF PCR Purification Kits utilize multiwell technology for manual or fully automated high-throughput purification, using an ultrafiltration-plate format. The purification procedure allows small molecules such as primers, salts, and unincorporated nucleotides to pass through the membrane of the ultrafiltration plate, while PCR products of >100 bp are retained.

Procedure

The MinElute and QIAquick Systems use a simple bindwash- elute procedure. Binding buffer is added directly to the PCR sample or other enzymatic reaction, and the mixture is applied to the MinElute or QIAquick spin column, or QIAquick 8-well strip or 96-well plate. Gel slices are dissolved in a buffer containing a pH indicator, allowing easy determination of the optimal pH for DNA binding. Nucleic acids adsorb to the silica-gel membrane in the high-salt conditions provided by the buffer. Impurities are washed away and pure DNA is eluted with a small volume of low-salt buffer provided or water, ready to use in all subsequent applications.

MinElute 96 UF PCR Purification Kits use ultrafiltration technology. PCR products are loaded into the wells of the ultrafiltration plates, and a vacuum is applied. While small molecules such as primers, salts, and unincorporated nucleotides pass through the membrane, PCR products >100 bp are retained. Purified PCR products are eluted directly from the surface of the membrane in small volumes (as little as 20 µl in the manual procedure or 30 µl on BioRobot workstations), leading to highly concentrated DNA eluates. Water, DMSO, or buffers such as SSC can be used for elution.

Handling

MinElute and QIAquick spin columns are designed to provide two convenient handling options (see flowcharts). The spin columns fit into a conventional table-top microcentrifuge or onto any vacuum manifold with luer connectors, such as QIAvac 24 or QIAvac 6S with QIAvac Luer Adapters (see figure "MinElute and QIAquick Spin Column Handling Options"). QIAquick multiwell modules are processed using vacuum-driven purification on QIAvac manifolds; QIAquick 8 Strips are used on QIAvac 6S, and QIAquick 96 Plates on QIAvac 96. The QIAquick 96 procedure can also be automated on BioRobot Systems.

MinElute and QIAquick Spin Column Handling Options


	Spin columns: A in a microcentrifuge, B on QIAvac Luer Adapters, C on other luer connectors.

Downstream applications

DNA fragments purified with the MinElute or QIAquick System are ready for direct use in all applications, including:

Radioactive and fluorescent sequencing
Microarray analysis
Ligation and transformation
Restriction digestion
Labeling
Microinjection
PCR
In vitro transcription