从细胞或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 Micro Kit可以从同一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柱流出液流过RNeasy MinElute离心柱纯化得到RNA。该试剂盒最多可处理5 x 105的细胞或5 mg的组织样本。如需处理更多样本,可使用AllPrep DNA/RNA Mini Kit。
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AllPrep DNA/RNA Micro Kit
For 50 micropreps: AllPrep DNA Spin Columns, RNeasy MinElute Spin Columns, Collection Tubes, Carrier RNA, RNase-Free Water and Buffers
80284
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The AllPrep DNA/RNA Micro 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.|Wide dynamic range.|
|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 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.|
Performance

AllPrep DNA/RNA Micro Kit可处理培养细胞或易裂解组织(见表)。应用新型AllPrep DNA离心柱纯化基因组DNA,RNeasy MinElute离心柱纯化总RNA。通常从培养细胞中纯化所得总RNA的RIN值接近10。纯化的基因组DNA的平均长度为15–30 kb,取决于匀浆情况(参见"Simultaneous purification of total RNA and genomic DNA")。可在很大范围内对基因组DNA和总RNA进行可靠的定量检测(参见"Wide dynamic range")。

从组织中纯化DNA和RNA的最佳产量
组织(5 mg) DNA产量(µg)* RNA产量(µg)*
心脏 3.89 2.00
大脑 2.60 3.08
11.33 12.70
32.40 16.02
9.65 24.63
20.50 43.51
胸腺 34.90 56.89
26.71 27.38
* DNA和RNA是从每种组织的2个5 mg样本中纯化得到,表中为平均产量。
Principle

AllPrep DNA/RNA Micro Kit用于从单个细胞或组织样本中纯化基因组DNA和总RNA,非常适合处理小样本,诸如激光显微切割的冻干切片和细针穿刺抽出液等。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40分钟内从同一样本中纯化高品质DNA和RNA(参见"AllPrep DNA/RNA procedure")。可处理培养细胞和易裂解组织,应用新型AllPrep DNA离心柱纯化基因组DNA,RNeasy MinElute离心柱纯化总RNA。

在Buffer RLT Plus(AllPrep DNA/RNA Micro 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。

另可选择补充操作流程纯化蛋白质。纯化所得的蛋白是变性的,可用于诸如SDS-PAGE、Western印迹和2D凝胶电泳等应用。对于更大样品量的实验,可使用AllPrep DNA/RNA/Protein Mini Kit从细胞或组织样品中同时纯化DNA、RNA和蛋白。

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。纯化所得的蛋白是变性的,可用于以下应用:SDS‑PAGE、Western印迹和2D凝胶电泳。

Feature
Specifications
Applications PCR, RT-PCR, real-time PCR, cDNA synthesis, microarray, blotting
Elution volume DNA: 30 µl; RNA: 10 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 and total RNA
Sample amount 5 x 10e5 cells or 5 mg tissue
Technology Silica technology
Time per run or per prep 35 min
Yield Varies

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FAQs
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Kit Handbooks
1
For simultaneous purification of genomic DNA and total RNA from the same small sample, including
  • animal and human cells (≤5 x 10^5)
  • animal and human tissues (≤5 mg)
  • microdissected cryosections

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