MinElute Reaction Cleanup Kit

For cleanup of up to 5 µg DNA (70 bp to 4 kb) from enzymatic reactions

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MinElute Reaction Cleanup Kit (50)

Cat. No. / ID: 28204

50 MinElute Spin Columns, Buffers, Collection Tubes (2&nbsp:ml)

DKK 1,205.00

Preparations

250

Quantity

The MinElute Reaction Cleanup Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

Very small elution volumes
Fast procedure and easy handling
High, reproducible recoveries
Gel loading dye for convenient sample analysis

Product Details

The MinElute Reaction Cleanup Kit provides spin columns, buffers, and collection tubes for silica membrane-based purification of DNA 70 bp – 4 kb in size from enzymatic reactions. The spin columns are designed to allow elution in very small volumes (as little as 10 µl), delivering highly concentrated DNA in high yields. An integrated pH indicator allows easy determination of the optimal pH for DNA binding to the spin column. The procedure can be fully automated on the QIAcube Connect. DNA fragments purified with the MinElute system are ready for direct use in all applications, including sequencing, microarray analysis, ligation and transformation, restriction digestion, labeling, microinjection, PCR, and in vitro transcription.

For optimal results it is recommended to use this product together with QIAvac 24 Plus.

Performance

The MinElute Reaction Cleanup Kit ensures cleanup of up to 5 µg DNA (70 bp to 4 kb) from enzymatic reactions, delivering high yields of DNA suitable for a range of applications. The kit provides spin columns for cleanup of enzymatic reactions. Using a microcentrifuge or vacuum manifold, high concentration of DNA fragment (70 bp – 4 kb) is quickly achieved. (DNA fragments larger than 4 kb should be purified using the QIAquick System.)

Examples of enzymes that are completely removed by the MinElute Reaction Cleanup Kit
Protein	Molecular weight per enzyme subunit (kDa)
DNA Polymerase I	109
Klenow fragment	62
Calf intestinal alkaline phosphatase	69
T4 DNA ligase	55
T4 Polynucleotide kinase	35
Terminal transferase	32
DNase I	31
Restriction enzymes	Varies

Principle

MinElute Kits contain a silica 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.

Gel loading dye

To enable faster and more convenient sample processing and analysis, gel loading dye is provided. GelPilot Loading Dye contains three tracking dyes (xylene cyanol, bromophenol blue, and orange G) to facilitate the optimization of agarose gel run time and prevent smaller DNA fragments migrating too far (see figure " GelPilot Loading Dye").

See figures

GelPilot Loading Dye.

Procedure

The MinElute system uses a simple bind-wash-elute procedure (see flowchart " MinElute procedure"). Binding buffer is added directly to the enzymatic reaction, and the mixture is applied to the MinElute spin column. The binding buffer contains a pH indicator, allowing easy determination of the optimal pH for DNA binding (see figure "pH Indicator Dye"). Nucleic acids adsorb to the silica 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 subsequent applications.

Handling

MinElute spin columns are designed to provide two convenient handling options (see flowchart "MinElute procedure"). The spin columns fit into a conventional table-top microcentrifuge or onto any vacuum manifold with luer connectors, such as QIAvac 24 Plus or QIAvac 6S with QIAvac Luer Adapters. The MinElute Reaction Cleanup Kit, in addition to other QIAGEN spin-column-based kits, can be fully automated on the QIAcube Connect, enabling increased productivity and standardization of results (see figures "Spin column handling options A, B, C, D, and E" and " QIAcube Connect").

See figures

MinElute procedure.

pH Indicator Dye.

Spin column handling options — A.

Spin column handling options — B.

Spin column handling options — C.

Spin column handling options — D.

Spin column handling options — E.

QIAcube Connect.

Applications

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

Sequencing
Microarray analysis
Ligation and transformation
Restriction digestion
Labeling

Supporting data and figures

Spin column handling options — E.

Specifications

Features	Specifications
Binding capacity	5 µg
Fragment size	70 bp – 4 kb
Elution volume	10 µl
Recovery: oligonucleotides dsDNA	Recovery: oligonucleotides, dsDNA
Removal <10mers 17–40mers dye terminator proteins	Removal <40mers
Format	Tube
Sample type: applications	DNA, oligonucleotides: Enzymatic reactions
Technology	Silica technology

Resources

Download Safety Data Sheets for QIAGEN product components.

FAQ

Buffer PB of the QIAquick PCR Purification Kit cannot be used to extract DNA from agarose gels. However, Buffer QG of the QIAquick Gel Extraction Kit can be used to remove salt and proteins from enzymatic reactions by adding 3 volumes of Buffer QG and 1 volume of isopropanol to the reaction and proceeding with step 6 of the Gel Extraction Spin Protocol in the QIAquick Spin Handbook. See the QIAquick Spin Handbook for a list of reactions which can be cleaned up with the various QIAquick kits.

FAQ ID -786

The MinElute spin columns included in the following kits should be stored at 2–8°C upon arrival: AllPrep DNA/RNA Micro, EpiTect Fast DNA Bisulfite, EpiTect Fast FFPE Bisulfite, EpiTect Fast LyseAll Bisulfite, EpiTect Plus DNA Bisulfite, EpiTect Plus FFPE Bisulfite, EpiTect Plus LyseAll Bisulfite, exoRNeasy Serum/plasma Maxi, exoRNeasy Serum/Plasma Midi, GeneRead DNA FFPE, GeneRead rRNA Depletion, GeneRead Size Selection, MinElute Gel Extraction, MinElute PCR Purification, MinElute Reaction Cleanup, miRNeasy FFPE, miRNeasy Micro, miRNeasy Serum/Plasma, QIAamp DNA FFPE, QIAamp DNA Investigator, QIAamp DNA Micro, QIAamp MinElute Media, QIAamp MinElute Virus Spin, QIAamp MinElute Virus Vacuum, RNeasy FFPE, RNeasy Micro, RNeasy Plus Micro.

Short-term storage (up to 4 weeks) at room temperature (15–25°C) does not affect the performance. However, for optimal performance and quality, storage temperature should not exceed 25°C.

FAQ ID - 3560

Yes, and therefore they are interchangeable.

FAQ ID -581

DNA fragments purified with the QIAGEN DNA Cleanup Systems, i.e., the QIAquick PCR Purification Kit, the MinElute Reaction Cleanup Kit, the QIAEX II Gel Extraction Kit etc. may float out of the loading wells of agarose gels due to residual ethanol carried over from the wash step with Buffer PE (despite the addtition of glycerol-containing loading buffer).

Use either of the following options to remove residual ethanol from the eluate:

re-purify the sample using a QIAquick-, or MinElute column, or QIAEX II resin
incubate the eluate at 56°C for 10 min to evaporate the ethanol
dry down the sample in a vacuum centrifuge, and resuspend the pellet in a small volume of sterile water

FAQ ID -205

As a rule of thumb, single-stranded DNA binds to silica with approximately half the affinity of a double-stranded DNA fragment of the same length under the buffer conditions used in the QIAquick and MinElute Kits. Even though no systematic experimental data exists, we expect that recovery of ssDNA fragments of approximately 200 nucleotides and below will not be very efficient after cleanup using the QIAquick PCR Purification Kit or MinElute PCR Purification Kit. By comparison, it should be possible to purify fragments longer than 140 nucleotides using the QIAquick Gel Extraction Kit.

Note that recovery of single strand DNA is influenced to some degree also by factors such as base composition and secondary structure. It has to be determined empirically by the researcher if cleanup of single-stranded DNA with QIAquick columns yields satisfactory results.

FAQ ID -759

The composition of Buffer EB is:

10 mM Tris-Cl, pH 8.5

Buffer EB is the elution buffer used in the QIAquick PCR, Gel Extraction, Nucleotide Removal Kits, and MinElute Kits for DNA cleanup, and the QIAprep Miniprep Kits for small-scale plasmid purification. The purified DNA can also be eluted in TE (10 mM Tris-Cl, 1 mM EDTA, pH 8.0), but the EDTA may inhibit subsequent enzymatic reactions.

FAQ ID -199

Occasionally, DNA fragments eluted from the silica matrix of QIAquick, MinElute or QIAEX II Kits will contain denatured single-stranded DNA (ssDNA), appearing as a smaller band on an analytical gel. Under certain conditions, chaotropic agents (present in all silica-based DNA purification methods) can denature DNA fragments. This is a rare event that may be influenced by sequence characteristics such as the presence of inverted repeats or A–T-rich stretches.

Because salt and buffering agents promote renaturation of DNA strands, the following tips are recommended:

use the eluted DNA to prepare your downstream enzymatic reaction, but omit the enzyme. Incubate the reaction mix at 95°C for 2 minutes to reanneal the ssDNA, and allow the tube to cool slowly to room temperature before adding the enzyme and proceeding
alternatively, the DNA can be eluted from the silica-gel membrane or resin in 10 mM Tris buffer containing 10 mM NaCl. However, the salt concentration of the eluate must then be taken into consideration in downstream applications.

FAQ ID -148

The QIAquick Spin Columns (100) (cat. no. 28115) in the QIAquick PCR Purification, Gel Extraction, Nucleotide Removal and PCR & Gel Cleanup kits are also sold separately from the kits.

The MinElute columns in the MinElute PCR Purification, Gel Extraction and Reaction Cleanup kits are not sold separately.

We always provide extra buffers in our kits so you can scale up reactions, add extra washes or allow for spillage.

FAQ ID -2460