AllPrep DNA/RNA/Protein Mini Kit

从细胞和组织中同时纯化DNA、RNA和蛋白质

S_0508_GEX_AP_cmyk

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AllPrep DNA/RNA/Protein Mini Kit (50)

Cat. No. / ID:   80004

For 50 preps: AllPrep DNA Spin Columns, RNeasy Spin Columns, Collection Tubes, RNase-Free Reagents and Buffers
AllPrep DNA/RNA/Protein Mini Kit 旨在用于分子生物学应用。该产品不能用于疾病诊断、预防和治疗。

✓ 全天候自动处理在线订单

✓ 博学专业的产品和技术支持

✓ 快速可靠的(再)订购

特点

  • 从珍稀样本中最大程度回收各种待分析物
  • 简短流畅的操作流程,节省时间
  • 各分析物来自同一样本,数据可比
  • 无需苯酚或丙酮等有机溶剂

产品详情

AllPrep DNA/RNA/Protein Mini Kit可从同一细胞或组织样本中同时纯化DNA、RNA和蛋白质。该试剂盒可与Allprotect Tissue Reagent配合使用,此试剂可迅速稳定组织样本中的DNA、RNA和蛋白质,无需冷冻即可长期保存。可使用TissueRuptor或TissueLyser高效破碎样本。

绩效

使用AllPrep DNA/RNA/Protein Mini Kit纯化得到的基因组DNA和总RNA在real-time PCR和real-time RT-PCR分析中表现出色(参见" Reliable real-time PCR and RT-PCR analyses")。使用该试剂盒纯化得到的总蛋白质适用于SDS-PAGE和Western印迹(参见" Reliable western blotting and SDS-PAGE")。
查看图表

原理

AllPrep DNA/RNA/Protein Mini Kit使系统生物学研究中样本的制备过程标准化。DNA、RNA和蛋白质为同一来源,减少了从不同样本中制备这些分析物时引起的差异。使用此试剂盒无需在纯化前将样本分为三份,因此DNA、RNA和蛋白质的产量较高。此外,纯化过程无需使用苯酚和丙酮等有机溶剂。

程序

细胞或组织样本首先在Buffer RLT中裂解并匀浆,使得DNA酶、RNA酶和蛋白酶立刻失活,确保可纯化得到完整的DNA、RNA和蛋白质。用AllPrep DNA离心柱过滤裂解液。高盐缓冲液流过离心柱,基因组DNA可高效的选择性结合到柱上。洗脱之后即可得到纯化的即用型DNA。

在AllPrep DNA离心柱的流出液中加入乙醇,形成适宜RNA结合的条件。样本上样到RNeasy离心柱,总RNA与膜结合,污染物被去除。然后用不含RNA酶的纯水进行洗脱,得到高品质RNA。

Buffer APP是新型的水溶性蛋白沉淀溶液。将其加入到RNeasy离心柱的流出液中,然后离心得到蛋白质沉淀。用适合的溶液溶解完整的总蛋白质,可即用于下游应用。此试剂盒包含Buffer ALO,该缓冲液用于溶解蛋白沉淀,不影响SDS-PAGE操作(参见" AllPrep DNA/RNA/Protein procedure")。

查看图表

应用

AllPrep DNA/RNA/Protein Mini Kit可从同一细胞或组织样本中同时纯化基因组DNA、总RNA和总蛋白质,因此使样本制备流程更加高效。该试剂盒纯化得到的总蛋白适用于SDS-PAGE和Western印迹。使用AllPrep DNA/RNA/Protein Mini Kit纯化得到的基因组DNA和总RNA在多种下游应用中表现出色,包括:

  • Real-time PCR分析
  • Real-time RT-PCR分析

辅助数据和图表

Specifications

FeaturesSpecifications
ApplicationsPCR, RT-PCR,real-time PCR, real-time RT-PCR, Southern-,Northern-,Western Blotting, Microarrays, Genotyping,1D and 2D gel electrophoresis
Main sample typeEasy-to-lyse tissue or cells
Elution volumeRNA: 30 - 50µl, DNA: 100 - 200µl
Purification of total RNA, miRNA, poly A+ mRNA, DNA or proteinTotal RNA, genomic DNA and western-grade proteins
FormatSpin column
Yieldvaries, depends on sample type
Time per run or per prep60 min
ProcessingManual (centrifugation)
TechnologySilica technology, Protein precipitation
Sample amountup to 30 mg tissue or 1 x 10e7 cells

资源

试剂盒操作手册 (3)
For simultaneous purification of genomic DNA, total RNA, and total protein from the same cell or tissue sample
快速启动实验方案 (1)
For simultaneous purification of genomic DNA, total RNA and total protein from the same cell or tissue sample
安全数据表 (1)
Download Safety Data Sheets for QIAGEN product components.
Safety Data Sheets (1)
Certificates of Analysis (1)

FAQ

Can the AllPrep DNA/RNA/Protein Mini Kit be used with fibrous or lipid tissues?

The AllPrep DNA/RNA/Protein Mini Kit is optimized for use with easy-to-lyse tissues. For fibrous (e.g., heart) and lipid tissues (e.g., brain), we recommend using a portion of the tissue sample for RNA purification with the RNeasy Fibrous Tissue Mini Kit and RNeasy Lipid Tissue Mini Kit, respectively.

 

 

 

FAQ ID -1579
What is the composition of Buffer ALO?
The detailed composition of Buffer ALO is confidential. Buffer ALO is part of the AllPrep DNA/RNA/ Protein Kit and functions as protein resuspension buffer. Buffer ALO is a Laemmli-related sample buffer for use in SDS-PAGE. In addition to other reagents, Buffer ALO comprises SDS, bromophenol blue, and dithiothreitol (DTT). DTT is added by the user prior the first use of the kit.
FAQ ID -2805
Does Buffer APP in the AllPrep DNA/RNA/Protein Mini Kit contain acetone?

No, Buffer APP in the AllPrep DNA/RNA/Protein Mini Kit is a novel aqueous protein precipitation solution and has some advantages over acetone with regard to recovery of small proteins (about 10 kDa and smaller).

 

 

FAQ ID -1576
What effect does homogenization have on DNA yield and integrity when using AllPrep DNA/RNA Kits?

Yield and integrity of genomic DNA depend on the disruption and homogenization method used with the Allprep DNA/RNA Kits.

Homogenization with the TissueRuptor (or other rotor–stator homogenizer, such as the Polytron) or the TissueLyser results in greater DNA fragmentation (depending on homogenization time and intensity). However, shorter DNA fragments are easier to elute, so DNA yields will be higher.

In contast, homogenization using the QIAshredder (or a syringe and needle) is more gentle, resulting in less DNA fragmentation. However, longer DNA fragments are more difficult to elute, so DNA yields may be lower.

 

FAQ ID -1751
What can be done if the protein pellet does not solubilize completely using the AllPrep DNA/RNA/Protein Mini Kit?

In the protein precipitation procedure of the AllPrep DNA/RNA/Protein protocol, be sure to dissolve the protein pellet completely in Buffer ALO or 1x SDS-PAGE sample buffer.

For easier dissolving of proteins, dissolve the pellet in 5% (w/v) SDS or 8 M urea. Commonly, there is insoluble material left even after vigorous shaking or heating. This is typical for all kinds of protein precipitation under heavily denaturing conditions.

Briefly centrifuge the sample and use the supernatant for downstream analysis.

 

FAQ ID -1580
What is the composition of Buffer APP?
The exact composition of Buffer APP is confidential. Buffer APP is a component of the AllPrep DNA/RNA Protein Kit and functions as a protein precipitation buffer, containing, among other components, zinc chloride.
FAQ ID -2804
Can acetone be used for precipitation of protein from Buffer RLT lysates generated with RNeasy Kits?

Yes, please follow the Supplementary Protocol Acetone precipitation of protein from Buffer RLT or Buffer RLT Plus lysates (RY22).

Important Note:

Do not use TCA to precipitate protein from Buffer RLT and Buffer RLT Plus lysates. These buffers contain guanidine thiocyanate, which can form highly reactive compounds when combined with acidic solutions.

 

For simultaneous purification of DNA, RNA, and protein from the same sample (either cultured cells or easy-to-lyse tissues), we recommend using the AllPrep DNA/RNA/Protein Mini Kit. This kit allows precipitation of protein from Buffer RLT lysates using a novel protein precipitation buffer, Buffer APP.

Please note that Buffer APP is not compatible with Buffer RLT Plus. Acetone should be used instead to precipitate protein from RLT Plus lysates.

 

FAQ ID -1164
Can tissue homogenized in Buffer RLT Plus instead of Buffer RLT be used with the AllPrep DNA/RNA/Protein Mini Kit?

No, Buffer APP for protein precipitation in the Allprep DNA/RNA/protein Mini Kit is not compatible with Buffer RLT Plus. Acetone can be used to precipitate proteins from samples homogenized in Buffer RLT Plus contained in the Allprep DNA/RNA Mini Kit and RNeasy Plus Mini Kit.  

 

FAQ ID -1577
Why does AllPrep DNA/RNA/Protein Mini kit use buffer RLT, but Allprep DNA/RNA Mini and Allprep DNA/RNA Micro kits use buffer RLT Plus? Are these buffers interchangeable?

Buffer RLT Plus contains detergents that are more ideal for binding of DNA to the AllPrep DNA mini spin columns.

But RLT Plus is not compatible with Buffer APP used to precipitate proteins in AllPrep DNA/RNA/Protein Mini kit. Therefore, we chose RLT instead of RLT Plus to get both DNA and proteins with this kit.

RLT lysates can also be used in AllPrep RNA/DNA kit or the AllPrep DNA/RNA Micro kit. DNA and RNA yields should be fine, but RNA may contain slightly higher levels of genomic DNA contamination.

FAQ ID - 3391
3323 - Do you have a protocol for the AllPrep DNA/RNA/Protein Mini Kit on the QIAcube?
A custom protocol can be requested through the online QIAcube protocol request page: <https://www.qiagen.com/qiacube/customized/protocoladd.aspx >.
FAQ ID - 3323