MinElute Reaction Cleanup Kit

从酶促反应物中回收多至5 μg DNA(70 bp到4 kb)

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MinElute Reaction Cleanup Kit (50)

Cat. No. / ID:  28204

50 MinElute Spin Columns, Buffers, Collection Tubes (2&nbsp:ml)
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50
250

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

  • 非常小的洗脱体积
  • 回收速度快,操作容易
  • 回收率高,重复性好
  • 含有凝胶上样染料,方便样本分析

Product Details

MinElute Reaction Cleanup Kit含有离心柱、缓冲液和收集管,基于硅胶膜技术,用于从酶促反应物中纯化70 bp到4 kb的DNA片段。特殊设计的离心柱可将DNA洗脱至非常小的体积(10 μl),获得高产、高度浓缩的DNA。通过内置的pH指示剂可容易确定DNA结合到离心柱的最佳pH值。实验可在QIAcube全自动核酸纯化仪上全自动运行。应用MinElute System纯化的DNA可即用于各种应用,包括测序、微阵列分析、连接和转化、限制性酶切、标记、显微注射、PCR和体外转录。

Performance

MinElute Reaction Cleanup Kit可从酶促反应物中纯化多达5 μg的DNA(70 bp–4 kb),获得的高产量DNA适用于各种应用。该试剂盒提供纯化酶促反应物所需的离心管。使用离心机或真空装置可快速获得高度浓缩的DNA片段(70 bp–4 kb)。(大于4 kb的DNA片段使用QIAquick System纯化。)

MinElute Reaction Cleanup Kit完全清除的酶举例
酶的分子量(kDa)
DNA聚合酶I 109
Klenow片段 62
小牛肠碱性磷酸酶 69
T4 DNA连接酶 55
T4聚核苷酸激酶 35
末端转移酶 32
DNase I 31
限制性内切酶 不定

Principle

MinElute Kits采用硅胶膜式纯化柱,在高盐条件下结合DNA,低盐或水可洗脱DNA。纯化过程去除引物、核苷酸、酶、矿物油、盐、琼脂糖、溴化乙锭和DNA样品中的其他杂质。硅胶膜技术避免了松散树脂和悬液状态的问题及不方便性。经优化的特制结合缓冲液,专用于选择性吸附特定大小范围内的DNA分子。

凝胶上样染料

为更快速、更方便地进行分析,提供上样染料。GelPilot Loading Dye含有3种示踪染料(xylene cyanol、bromophenol blue和orange G),便于优化凝胶运行时间,避免小片段DNA跑得过远(参见" GelPilot Loading Dye")。

See figures

Procedure

MinElute体系应用简单的结合-洗涤-洗脱步骤(参见" MinElute procedure")。直接将结合缓冲液加入PCR样品或其他酶反应中,然后将混合液装载到MinElute离心柱上。结合缓冲液含有pH指示剂,可方便地确定DNA结合的最佳pH值(参见 pH Indicator Dye)。在高盐条件下,核酸吸附在硅胶膜上。洗去杂质并用少量的低盐缓冲液或水洗脱DNA,可即用于各种下游应用。

操作

MinElute离心柱有两种方便的处理方式(参见" MinElute procedure")。可可将离心柱放入传统的微型离心机或通过适配器连接到任何真空装置上,诸如通过QIAvac Luer Adapters连接到QIAvac 24 Plus或QIAvac 6S,也可在QIAcube全自动核酸纯化仪上全自动进行,确保高产量和标准化结果。

See figures

Applications

MinElute系列产品纯化的DNA片段可即用于各种应用,包括:

  • 测序
  • 微阵列分析
  • 连接和转化
  • 限制性酶切
  • 标记

Supporting data and figures

Resources

安全数据表 (1)
Download Safety Data Sheets for QIAGEN product components.
试剂盒操作手册 (1)
MinElute Handbook
PDF (611KB)
快速启动实验方案 (1)
Certificates of Analysis (1)
Kit Handbooks (1)
MinElute Handbook
PDF (611KB)
Quick-Start Protocols (1)

Publications

Oncocytic change in pleomorphic adenoma: molecular evidence in support of an origin in neoplastic cells.
Di Palma S; Lambros MB; Savage K; Jones C; Mackay A; Dexter T; Iravani M; Fenwick K; Ashworth A; Reis-Filho JS;
J Clin Pathol; 2006; 60 (5):492-9 2006 Feb 7 PMID:16467165
Similarity and differences in the Lactobacillus acidophilus group identified by polyphasic analysis and comparative genomics.
Berger B; Pridmore RD; Barretto C; Delmas-Julien F; Schreiber K; Arigoni F; Brüssow H;
J Bacteriol; 2006; 189 (4):1311-21 2006 Dec 1 PMID:17142402
Chromosome Conformation Capture Carbon Copy (5C): a massively parallel solution for mapping interactions between genomic elements.
Dostie J; Richmond TA; Arnaout RA; Selzer RR; Lee WL; Honan TA; Rubio ED; Krumm A; Lamb J; Nusbaum C; Green RD; Dekker J;
Genome Res; 2006; 16 (10):1299-309 2006 Sep 5 PMID:16954542
Quantitative proteomics of the archaeon Methanococcus maripaludis validated by microarray analysis and real time PCR.
Xia Q; Hendrickson EL; Zhang Y; Wang T; Taub F; Moore BC; Porat I; Whitman WB; Hackett M; Leigh JA;
Mol Cell Proteomics; 2006; 5 (5):868-81 2006 Feb 17 PMID:16489187
RAISE: a simple and novel method of generating random insertion and deletion mutations.
Fujii R; Kitaoka M; Hayashi K;
Nucleic Acids Res; 2006; 34 (4):e30 2006 Feb 21 PMID:16493137

FAQ

Are Buffer PB of the QIAquick PCR Purification Kit and Buffer QG of the QIAquick Gel Extraction Kit interchangeable?

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
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
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
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