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Nucleic Acid Purification


Products >  Nucleic_Acid_Purification >  RNA >  RNA-DNA-Protein >  RNA-DNA-Protein

Simultaneous Isolation of RNA, DNA, and Protein from One Lysate—NucleoSpin TriPrep

The NucleoSpin TriPrep kit allows you to isolate RNA, DNA, and protein simultaneously from one lysate, allowing sensitive gene expression analysis of small, rare, or precious samples. Parallel RNA, DNA, and protein extraction from the same experimental sample is increasingly in demand by researchers interested in gene regulation mechanisms, such as knockdown of mRNA expression by siRNAs and other processes that influence protein expression. This method is preferable to isolating RNA, DNA, and protein from different sample aliquots, which is difficult when the amount of starting material is limited, and necessitates proof of sample equality to provide reliable results.

The NucleoSpin TriPrep kit yields RNA, DNA, and protein in separate fractions from a single lysate in one working procedure without splitting the sample prior to extraction. Samples are lysed by incubation in a buffer containing large amounts of chaotropic salts, which inactivates enzymes such as RNases, phosphatases, and proteases that can degrade nucleic acids and proteins. This lysis buffer thoroughly dissolves even barely soluble proteins, and also provides appropriate binding conditions for RNA and DNA. Thus, nucleic acids and protein from the same sample can be separated by simply centrifuging the lysate through the NucleoSpin TriPrep spin column: RNA and DNA are bound to the silica membrane while the protein is recovered in the column flowthrough (see procedure).

The nucleic acids are separated by sequential elution steps. First the DNA is eluted with DNA Elute, a low ionic strength buffer, while the RNA is still bound to the column. DNA elution is followed by wash steps, on-column DNA digestion (to remove residual DNA contamination), and elution of high-quality RNA. Proteins in the flowthrough are precipitated by a special buffer (Protein Precipitator), pelleted by centrifugation, washed, and then dissolved in Protein Solving Buffer. Isolated RNA and DNA are of high quality and ready to use for all common downstream applications. Isolated protein may be used directly for quantification, SDS-PAGE, and Western blot analysis.

  At-A-Glance   Documents   Images & Data

Features

  • Convenient single-column isolation of RNA, DNA, and protein
  • Provides high-quality RNA and DNA, ready to use for all typical downstream applications
  • High protein yield is independent of protein size, localization, modification, etc.
  • Complete kit includes shredders, on-column rDNAse, and a buffer for dissolving a variety of proteins
  • Preserves protein primary structure and post-translational modifications (e.g., protein phosphorylation), allowing protein analysis without additional inhibitors (e.g., proteinase or phosphatase inhibitors)

Applications

  • Rapid purification of total RNA, DNA, and protein from small and precious samples—without splitting samples
  • Gene expression profiling, analysis of transgenic organisms, drug screening, genotyping
  • Reliable interpretation of RNA, DNA, and protein amounts
  • Isolation of RNA and DNA that is ready to use for all typical downstream applications
  • RNA, DNA and protein purification from a broad spectrum of starting materials, including cultured cells, tissue, bacteria, yeast, and plants
  • Purification of proteins that are ready to use for SDS-PAGE/Western blotting

Product Overview

Technology Silica membrane technology
Format Mini spin columns
Starting Material <5 x 106 cultured cells,
 <30 mg human/animal tissue,
 <100 mg plant tissue

RNA

DNA

Protein

Fragment Size

>200 nt

<30 kb

15–300 kb

Typical Yield

<70 µg

<6 µg

<1,200 µg

A260/280

1.9–2.1

1.7–1.9

Typical RIN (RNA integrity number)

>9

Elution Volume (RNA and DNA)
Resolubilization Volume (protein)

40–120 µl

100 µl

10–100 µl

Preparation Time

30 min/60 preps

35 min/6 preps (RNA + DNA)

30 min/6 preps

Binding Capacity

200 µg

10 µg*

* The binding capacity for DNA is <10 μg, and depends strongly on the amount of RNA bound to the membrane.


 
 
Products
Offers Cat. # Product Contents Size Price Units Select
740966.50 NucleoSpin® TriPrep 50 Preps $405.00
740966.250 NucleoSpin® TriPrep 250 Preps $1,816.00
 

 

 
 

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