MinElute PCR Purification Kit

For purification of up to 5 μg PCR products (70 bp to 4 kb) in low elution volumes

Features

  • Very small elution volumes
  • Fast procedure and easy handling
  • High, reproducible recoveries
  • Gel loading dye for convenient sample analysis
MinElute PCR Purification Kit (1000)

Cat. No. / ID: 28006X4

1000 MinElute Spin Columns, Buffers, Collection Tubes (2 ml)
€1,870.00
Preparations
1000
50
250
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The MinElute PCR Purification Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.

Product Details

The MinElute PCR Purification Kit provides spin columns, buffers, and collection tubes for silica-membrane-based purification of PCR products 70 bp – 4 kb in size. 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 optional 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.

Performance

The MinElute PCR Purification procedure removes primers, nucleotides, enzymes, mineral oil, salts, and other impurities from DNA samples (see figure " Efficient primer removal").

The MinElute PCR Purification Kit provides spin columns for PCR product cleanup. 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 PCR Purification Kit.)

See figures

Principle

The MinElute PCR Purification Kit contains 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. 

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

Procedure

The MinElute System uses a simple bind-wash-elute procedure. Binding buffer is added directly to the PCR sample or other 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 and can also be fully automated on the QIAcube (see figures "Spin column handling options  A,  B,  C,  D, and  E").

See figures

Applications

DNA fragments purified with the MinElute 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

Specifications

FeaturesSpecifications
bindingcapacity5 µg
sampletypeapplicationsDNA, oligonucleotides: PCR reactions
elutionvolume10 µl
fragmentsize70 bp – 4 kb
recoveryoligonucleotidesdsdnaRecovery: oligonucleotides, dsDNA
formatTube
technologySilica technology
processingManual
removal10mers1740mersdyeterminatorproteinsRemoval <40mers

Publications

Factors involved in root formation in Medicago truncatula.
Imin N; Nizamidin M; Wu T; Rolfe BG;
J Exp Bot; 2006; 58 (3):439-51 2006 Dec 6 PMID:17158109
Expression of c-kit in human osteosarcoma and its relevance as a prognostic marker.
Sulzbacher I; Birner P; Toma C; Wick N; Mazal PR;
J Clin Pathol; 2006; 60 (7):804-7 2006 Oct 3 PMID:17018686
Application of microdroplet PCR for large-scale targeted bisulfite sequencing.
Komori HK; LaMere SA; Torkamani A; Hart GT; Kotsopoulos S; Warner J; Samuels ML; Olson J; Head SR; Ordoukhanian P; Lee PL; Link DR; Salomon DR;
Genome Res; 2011; 21 (10):1738-45 2011 Jul 14 PMID:21757609
Transcriptional organization, regulation and role of the Porphyromonas gingivalis W83 hmu haemin-uptake locus.
Lewis JP; Plata K; Yu F; Rosato A; Anaya C;
Microbiology (Reading); 2006; 152 (Pt 11):3367-3382 2006 Nov PMID:17074906

FAQ

I received a kit containing the MinElute columns; however, they were left out for a while and not stored at 2–8°C upon receipt. Can I still use them?

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
Do CoralLoad dyes supplied in various QIAGEN PCR Kits interfere with downstream applications?

CoralLoad dyes supplied in PCR Kits such as, e.g., Taq, HotStarTaq, and TopTaq DNA Polymerase and TopTaq Master Mix do not interfere with most downstream enzymatic applications.

However, for reproducible results, purification of PCR products using the QIAquick or MinElute PCR Purification Kits prior to enzymatic manipulation is recommended.

 

 

FAQ ID -1745
What is the composition of Buffer PB?
Buffer PB contains a high concentration of guanidine hydrochloride and isopropanol. The exact composition of Buffer PB is confidential. However, this buffer can be purchased separately: Buffer PB.
FAQ ID -2791
Do I have to remove the oil from my PCR reaction before using the QIAquick or MinElute PCR Purification Kit?
No - mineral oil will not affect the clean-up procedure with the QIAquick or MinElute PCR Purification Kit.
FAQ ID -575
Are the columns of the MinElute Reaction Cleanup-, Gel Extraction-, and PCR Purification Kit identical?
Yes, and therefore they are interchangeable.
FAQ ID -581
Why does my DNA sample float out of the slot when loading it onto an agarose gel?

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
Do you have information about the cleanup of single-stranded DNA (ssDNA) with QIAquick columns?

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
What is the composition of Buffer EB?

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
Do you have a forensic post-PCR purification protocol to purify double-stranded DNA fragments from PCR reactions?

Yes, please follow the User-developed protocol 'Forensic post-PCR purification protocol using the MinElute PCR Purification Kit' (ME01).

 

 

 

FAQ ID -1761
What is the small band below my fragment of interest on an agarose gel after DNA cleanup using QIAquick?

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
Can I buy QIAquick and MinElute columns separately?

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