从细胞和组织样本中同时纯化DNA和RNA

  • 从同一样本中获得高品质DNA和RNA
  • 从珍稀样本中得到较大量的DNA和RNA
  • 简短流畅的操作流程,实现快速纯化
  • 即用型DNA和RNA,用于多种下游应用
AllPrep DNA/RNA Mini Kit可以从同一个生物样本中同时纯化基因组DNA和总RNA。通过新型的AllPrep DNA离心柱纯化DNA,然后AllPrep柱流出液经过RNeasy MinElute离心柱纯化得到RNA。该试剂盒可在QIAcube全自动核酸纯化仪上自动运行,最多可处理107个细胞或30 mg的组织样本。
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AllPrep DNA/RNA Mini Kit (50)
For 50 minipreps: AllPrep DNA Spin Columns, RNeasy Mini Spin Columns, Collection Tubes, RNase-Free Water and Buffers
80204
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The AllPrep DNA/RNA Mini Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
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AllPrep DNA/RNA procedure.|Simultaneous purification of total RNA and genomic DNA.|Multiplex PCR of 8 targets.|Wide dynamic range.|High-quality total RNA.|
|Total RNA and genomic DNA were purified from 10 mg samples of rat liver using AllPrep technology (the AllPrep DNA/RNA Mini Kit). [A] Purified RNA samples were analyzed on a 1.2% formaldehyde agarose gel. [B] Purified DNA samples were analyzed on a 0.8% agarose gel. M: markers.|Genomic DNA was purified from mouse tissues using the AllPrep DNA/RNA Mini Kit. Multiplex PCR of 8 targets (192–1953 bp; GC content in the range of 19–61%) was performed using 100 ng DNA per sample. M: markers; NTC: no template control; Li: liver; K: kidney; Lu: lung.|Genomic DNA and total RNA were purified from 10-fold serial dilutions of rat kidney homogenate using the AllPrep DNA/RNA Micro Kit; at each dilution, DNA was purified from duplicates, and RNA was purified from 4 replicates. [A] Real-time PCR analysis of the Pgk1 gene and [B] real-time RT-PCR analysis of the Jun transcript showed reliable quantification over a wide dynamic range, from 5 mg down to 50 ng tissue.|Total RNA was purified from 1 x 106 Jurkat cells using the AllPrep DNA/RNA Mini Kit and analyzed on the Agilent 2100 Bioanalyzer. The RIN value was 10, indicating intact RNA. M: marker; 18S: 18S rRNA; 28S: 28S rRNA.|
Performance
简单的流程可从同一样本中纯化高品质DNA和RNA。纯化得到的基因组DNA的平均长度为15–30 kb,适用于多重PCR(参见"Multiplex PCR of 8 targets")。纯化得到高品质的总RNA,RIN值接近10,表明RNA是完整的(参见"Simultaneous purification of total RNA and genomic DNA"和"High-quality total RNA")。AllPrep DNA/RNA纯化技术纯化得到的基因组DNA和总RNA在下游分析中可获得可靠的定量结果(参见"Wide dynamic range")。
Principle

AllPrep DNA/RNA Mini Kit用于从单个细胞或组织样本中纯化基因组DNA和总RNA。由于无需将样本分为两份单独进行纯化,可获得较大产量的DNA和RNA。经纯化的DNA和RNA可分别洗脱并直接用于多种下游应用。

Procedure

简单的流程可在35分钟内从同一样本中纯化高品质DNA和RNA(参见"AllPrep DNA/RNA procedure")。培养细胞和易裂解组织均可处理,应用新型AllPrep DNA离心柱纯化基因组DNA,RNeasy MinElute离心柱纯化总RNA。另可选择补充操作流程纯化蛋白质。纯化所得蛋白是变性的,可用于诸如SDS-PAGE、Western印迹和2D凝胶电泳等应用。对于经常从单一样本中纯化DNA、RNA和蛋白质的实验,可使用AllPrep DNA/RNA/Protein Mini Kit。

在Buffer RLT Plus(AllPrep DNA/RNA Mini Kit中提供)中破碎并匀浆组织时,可能出现很多泡沫。可在开始破碎和匀浆前,在Buffer RLT Plus加入Reagent DX(单独订购),终浓度为0.5%(v/v),即可有效消除泡沫。Reagent DX已经过精密测试,对于RNA纯度和诸如real-time RT-PCR等下游应用没有影响。添加了Reagent DX的Buffer RLT Plus可在室温(15–25ºC)下至少保存9个月。

Applications

纯化的基因组DNA的平均长度为15–30 kb,取决于匀浆情况,适合多种应用,包括:Southern印迹、斑点杂交和狭缝印迹、DNA甲基化分析、PCR和多重PCR。

经纯化的总RNA可即用于多种应用,包括:RT‑PCR和real‑time RT‑PCR、差异显示、cDNA合成、Northern印迹、斑点杂交和狭缝印迹、微阵列分析。

另可选择补充操作流程纯化蛋白质。纯化所得蛋白是变性的,可用于以下应用:SDS-PAGE 、Western印迹、2D凝胶电泳。

Feature
Specifications
Applications PCR, real-time PCR, microarray, blotting
Elution volume 100 µl
Format Spin column
Main sample type Cells, tissue
Processing Manual (centrifugation)
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein Genomic DNA, total RNA
Sample amount 1 x 10e7/30 mg
Technology Silica technology
Time per run or per prep 35 minutes
Yield Varies

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Kit Handbooks
1
For simultaneous purification of genomic DNA and total RNA from the same animal cell or tissue sample
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