MinElute Gel Extraction Kit

For gel extraction of up to 5 µg DNA fragments (70 bp to 4 kb) in low elution volumes

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

MinElute Gel Extraction Kit (50)

Cat. No. / ID: 28604

50 MinElute Spin Columns, Buffers, Collection Tubes (2 mL)

Preparations

250

The MinElute Gel Extraction 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 Gel Extraction Kit provides spin columns, buffers, and collection tubes for silica-membrane-based purification of DNA fragments of 70 bp – 4 kb from up to 400 mg gel slices. The spin columns are designed to allow elution in very small volumes (as little as 10 µL), delivering high yields of highly concentrated DNA. An integrated pH indicator allows easy determination of the optimal pH for DNA binding to the spin column. 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. The MinElute Gel Extraction Kit can be automated on the QIAcube Connect.

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

Performance

The MinElute Gel Extraction Kit removes nucleotides, enzymes, salts, agarose, ethidium bromide, and other impurities from DNA samples, delivering highly concentrated DNA suitable for a variety of downstream applications (see figure " Higher DNA concentrations").

The MinElute Gel Extraction Kit provides spin columns for gel extraction. Using a microcentrifuge or vacuum manifold, high concentration DNA of 70 bp – 4 kb is quickly purified. (DNA fragments from 4 kb – 10 kb should be purified using the QIAquick Gel Extraction Kit and DNA fragments smaller than 70 bp or larger than 10 kb should be extracted with the QIAEX II Gel Extraction System.)

See figures

Higher DNA concentrations.

Principle

MinElute Kits contain a silica membrane assembly for binding of DNA in high-salt buffer and elution with low-salt buffer or water. 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"). Gel slices are dissolved in a buffer containing a pH indicator, allowing easy determination of the optimal pH for DNA binding (see figure " pH Indicator Dye"), and the mixture is applied to the MinElute spin column. 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 subsequent applications.

Handling

MinElute spin columns are designed to provide two convenient handling options. The spin columns fit into a conventional table-top microcentrifuge or onto any vacuum manifold with luer connectors, such as the QIAvac 24 Plus with QIAvac Luer Adapters. The MinElute Gel Extraction 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 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.

QIAcube Connect.

Applications

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

Sequencing, including next generation sequencing
Microarray analysis
Ligation and transformation
Restriction digestion
Labeling

Supporting data and figures

Spin column handling options — C.

Manifold with luer connectors

Specifications

Features	Specifications
Binding capacity	5 µg
Elution volume	10 µL
Fragment size	70 bp – 4 kb
Sample type: applications	DNA: PCR reactions
Technology	Gel extraction
Recovery: oligonucleotides dsDNA	Recovery: dsDNA fragments
Format	Tube
Processing	Manual

Resources

Publications

MicroRNA-137 targets microphthalmia-associated transcription factor in melanoma cell lines.

Bemis LT; Chen R; Amato CM; Classen EH; Robinson SE; Coffey DG; Erickson PF; Shellman YG; Robinson WA;

Cancer Res; 2008; 68 (5):1362-8 2008 Mar 1 PMID:18316599

Molecular and phylogenetic analyses reveal mammalian-like clockwork in the honey bee (Apis mellifera) and shed new light on the molecular evolution of the circadian clock.

Rubin EB; Shemesh Y; Cohen M; Elgavish S; Robertson HM; Bloch G;

Genome Res; 2006; 16 (11):1352-65 2006 Oct 25 PMID:17065608

An accurate fluorescent assay for quantifying the extent of RNA editing.

Roberson LM; Rosenthal JJ;

RNA; 2006; 12 (10):1907-12 2006 Sep 6 PMID:16957279

Adaptive evolution of fertilization proteins within a genus: variation in ZP2 and ZP3 in deer mice (Peromyscus).

Turner LM; Hoekstra HE;

Mol Biol Evol; 2006; 23 (9):1656-69 2006 Jun 14 PMID:16774977

Collision events between RNA polymerases in convergent transcription studied by atomic force microscopy.

Crampton N; Bonass WA; Kirkham J; Rivetti C; Thomson NH;

Nucleic Acids Res; 2006; 34 (19):5416-25 2006 Sep 29 PMID:17012275

FAQ

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

Cut out the slice of agarose containing the DNA fragment of interest, and store it at 4^oC in an Eppendorf tube sealed with Parafilm.

FAQ ID -313

Yes, and therefore they are interchangeable.

FAQ ID -581

Yes. The QIAquick Gel Extraction Kit for extraction of DNA from gels can also be used for RNA gel extraction. Please see a user-developed procedure below, which was kindly provided by J. Knobloch, Heinrich Heine University, Düsseldorf, Germany. Note that this protocol has not been thoroughly tested and optimized by QIAGEN. QIAquick Gel Extraction Kits are not guaranteed to be RNase-free.

Excise the RNA fragment from the formaldehyde agarose gel with a clean, sharp scalpel.
Weigh the gel slice, and record the weight. Soak the gel slice in TE buffer for 25 min at room temperature with gentle shaking.
Remove the gel slice from the TE buffer, and place it in a colorless tube. Add 6 volumes of Buffer QG to 1 volume of gel, based on the gel weight (100 mg ~ 100 µl).
Incubate at 58°C for 25 min. To help dissolve the gel, mix by vortexing the tube every 2–3 min during the incubation.
Continue the QIAquick Gel Extraction Kit Protocol (using a microcentrifuge) in the QIAquick Spin Handbook, beginning with step 4.

Please click here for Figure 1.

Figure 1:

A. Total RNA was isolated from the parasitic blood fluke Schistosoma mansoni using the RNeasy Mini Kit and run on a formaldehyde agarose (1.2%) gel. (Note: The 28S rRNA in S. mansoni contains a break site so that the rRNA splits into two parts, which run on a gel at the same size as the 18S rRNA.) The rRNA bands were excised and treated as described above (left lane) or using 10 volumes of Buffer QG in step 3 (right lane).

B. The extracted RNA was then analyzed on a new formaldehyde agarose gel. (Data kindly provided by J. Knobloch, Department of Genetic Parasitology, Heinrich Heine University, Düsseldorf, Germany).

FAQ ID -133

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

The QIAEX II and QIAquick Gel Extraction Kit can be used to extract DNA from polyacrylamide gels.

The QIAEX II Handbook contains a protocol for Polyacrylamide Gel Extraction. A specialized User-Developed Protocol (QQ05) is available when using the QIAquick Gel Extraction Kit for this purpose.

Both protocols require the preparation of a diffusion buffer and a disposable plastic column or syringe barrel containing a Whatman GF/C filter or siliconized glass wool. To ensure optimal diffusion, cut the gel slices as small as possible, and use 2 volumes of diffusion buffer per 1 volume of gel. Increasing incubation time (protocol step 3) may result in higher yields.

FAQ ID -120