This website would like to remind you: Your browser (IE8) is out of date.Update your browser for more security, comfort and the best experience on this site.
×
Quote Request

RNA Isolation

Isolation from Tissue

Agencourt RNAdvance Tissue

The Agencourt RNAdvance Tissue total RNA extraction technology in combination with Beckman Coulter automation offers researchers a true walk away solution for consistent recovery of high quality total RNA in a multi-well format. Using Solid Phase Reversible Immobilization (SPRI) paramagnetic bead-based technology, the Agencourt RNAdvance Tissue system can extract total RNA from a wide variety of tissues without the hazards and waste removal issues of organic solvents. The Agencourt RNAdvance Tissue system produces high recovery of total RNA from soft, fibrous and lipid-rich tissues.

Overview

  • Extraction and purification of high quality total RNA from many tissue types
  • Efficient removal of genomic DNA and other contaminants
  • No centrifugation or vacuum filtration required
  • No organic extraction steps
  • Supports automated as well as manual processing
  • Ability to process from eight to 96 samples on a Biomek NX Workstation with a Span-8 configuration


Application: Total RNA Extraction From Tissue
Downstream Application Techniques: qRT-PCR, Microarray Analysis


Process Overview

Manual Process: A. Add tissue to lysis buffer B. Mechanically homogenize and PK digestion.
Automated or Continued Manual Process: 1. Add Binding Buffer 2. Separate magnetic beads from supernatant,
wash with Ethanol 3. DNase I (Optional) 4. Add Rebinding Buffer (Only required when optional DNase
step is performed) 5. Separate magnetic beads from supernatant, wash with Ethanol 6. Elute


High Yield

Isolating total RNA using the Agencourt RNAdvance Tissue protocol results in high yields without the necessity of centrifugation, vacuum filtration or organic solvents. Isolation of RNA from 10 mg of rat liver using the Agencourt RNAdvance Tissue process resulted in greater than three-fold increase in yield over RNeasy* and greater than two-fold increase in yield over MagMax* (Figure 1). As shown in Figure 2, the Agencourt RNAdvance Tissue system produced a lower Ct value indicating an increased starting quantity of RNA.

 
Figure 1                                                       Figure 2

 
 
Figure 1. The Agencourt RNAdvance Tissue system was compared to competitor kits by extracting 10 mg of rat liver tissue in a multi-well format, N = 24 for each kit.


Figure 2. Equal amounts of rat liver was processed using Agencourt RNAdvance and a competitor RNA extraction kit for 8 replicates each. The RT reaction was performed using the Invitrogen first strand synthesis kit using an equal amount from each kit. The qPCR reaction was performed using Applied Biosystems TaqMan Universal PCR Master Mix* on an ABI Prism* 7900. The primers and probe were designed to amplify the rat beta-actin gene.









 


Variety of Tissue Types


The ability to isolate total RNA can vary greatly between tissue types due to many factors including differences in endogenous levels of RNases and the fibrous or lipid-rich nature of certain tissues. The Agencourt RNAdvance Tissue procedure has been shown to work well with a wide variety of tissue samples. Figure 3 shows the yield of RNA obtained from extraction of five different tissue types including fibrous and fatty tissues.




Figure 3. Total RNA was extracted from several different rat tissues sing the Agencourt RNAdvance Tissue system on a Biomek NX Span-8. Yields are reported as µg/mg of tissue.
N = 24 for liver tissue, N = 8 for all other tissues.


Quality RNA Purification
 

RNA quality can be readily determined by electrophoresis on an Agilent* Bioanalyzer 2100. Intact, high-quality RNA will have well-defined 28s and 18s peaks with the absence of peak shoulders or an elevated baseline, which are both indicative of degradation. The Bioanalyzer software can calculate an RNA Integrity Number (RIN) based upon these and other qualities in the electropherogram. As seen in Figure 4, RNA extracted using the Agencourt RNAdvance Tissue system results in high RIN scores.

Figure 4. Agilent Bioanalyzer traces of RNA prepared from several
tissues using the Agencourt RNAdvance Tissue system


No Genomic DNA
 

To test for the presence of genomic DNA contamination, total RNA obtained from the Agencourt RNAdvance Tissue protocol was split into two reactions: one with reverse transcriptase and one without. The resulting samples were then put through PCR for the beta actin gene. The absence of an amplification product in the RT negative control (Figure 5) confirms that genomic DNA was not present in the total RNA.

 

Figure 5. One (1) µL of total RNA isolated from 10 mg rat liver was used as template for PCR. A 10 µl sample of the reactions were electrophoresed on 1.2% agarose gels. Top gel image: lanes 1-10: RT positive reactions. Lane 11: no template control. Bottom gel image: lanes 1-10: RT negative reactions. Lane 11: no template control. (PCR: amplicon is a 400bp region of the beta actin gene, 35 cycles)

Webinars & Symposiums

  • High-Performance RNA & miRNA Extractions using SPRI

    Bee Na Lee, PhD, Applications Scientist Beckman Coulter, Inc.

    Click to play  

  • Products

    Flow Cytometry Instruments & Software - Clinical Diagnostics

    Show: 25 50 100 items per page.

    Info
    Part No.
    Name
    Lasers
    Detectors
    Reg
    Price
    Buy

    Flow Cytometry Instruments & Software - Research and Discovery

    Show: 25 50 100 items per page.

    Info
    Part No.
    Name
    Lasers
    Detectors
    Reg
    Price
    Buy

    Centrifuges - Clinical Diagnostics

    Show: 25 50 100 items per page.

    Info
    Part No.
    Name
    Max Speed
    gForce
    Price
    Buy

    Centrifuges

    Show: 25 50 100 items per page.

    Info
    Part No.
    Name
    Max Speed
    gForce
    Price
    Buy

    Adapters - Clinical Diagnostics

    Show: 25 50 100 items per page.

    Info
    Part No.
    Name
    Material
    Dimensions
    Price
    Buy

    Adapters

    Show: 25 50 100 items per page.

    Info
    Part No.
    Name
    Material
    Dimensions
    Price
    Buy

    Nucleic Acid - Clinical Diagnostics

    Show: 25 50 100 items per page.

    Info
    Part No.
    Name
    Selection Options
    Sample Types
    Volume
    Reg
    Price
    Buy

    Nucleic Acid Sample Preparation

    Show: 25 50 100 items per page.

    Info
    Part No.
    Name
    Selection Options
    Sample Types
    Volume
    Reg
    Price
    Buy

    A32646

    Agencourt RNAdvance Tissue Kit - Large (384 preps)

    384 preps

    kr10 518,00

    A32649

    Agencourt RNAdvance Tissue Kit - Medium (96 preps)

    96 preps

    kr3 906,00

    A32645

    Agencourt RNAdvance Tissue Kit - Small (50 preps)

    50 preps

    kr2 550,00

    Showing 1 to 3 of 3 Results

    Liquid Handling Accessories - Clinical Diagnostics

    Show: 25 50 100 items per page.

    Info
    Part No.
    Name
    Mercury Content
    Humidity
    Operating Temp Range
    Price
    Buy

    Liquid Handling Accessories

    Show: 25 50 100 items per page.

    Info
    Part No.
    Name
    Mercury Content
    Humidity
    Operating Humidity
    Operating Temp Range
    Price
    Buy

    Liquid Handling Instruments - Clinical Diagnostics

    Show: 25 50 100 items per page.

    Info
    Part No.
    Name
    Air Supply Reqs
    Base Unit Depth
    Humidity
    Temp Range
    Price
    Buy

    Liquid Handlers

    Show: 25 50 100 items per page.

    Info
    Part No.
    Name
    Air Supply Reqs
    Base Unit Depth
    Humidity
    Temp Range
    Price
    Buy

    Liquid Handling Plates - Clinical Diagnostics

    Show: 25 50 100 items per page.

    Info
    Part No.
    Name
    Color
    Material
    Packaging
    Volume
    Well Shape
    Price
    Buy

    Liquid Handling Plates

    Show: 25 50 100 items per page.

    Info
    Part No.
    Name
    Color
    Material
    Packaging
    Volume
    Well Shape
    Price
    Buy

    Liquid Handling Reservoirs - Clinical Diagnostics

    Show: 25 50 100 items per page.

    Info
    Part No.
    Name
    Reservoir Material
    Tubing
    Divisions
    Material
    Packaging
    Volume
    Price
    Buy

    Liquid Handling Reservoirs

    Show: 25 50 100 items per page.

    Info
    Part No.
    Name
    Reservoir Material
    Tubing
    Divisions
    Material
    Packaging
    Volume
    Price
    Buy

    Liquid Handling Tips - Clinical Diagnostics

    Show: 25 50 100 items per page.

    Info
    Part No.
    Name
    Capacity (Max Vol)
    Packaging
    Reg
    Price
    Buy

    Liquid Handling Tips

    Show: 25 50 100 items per page.

    Info
    Part No.
    Name
    Capacity (Max Vol)
    Packaging
    Reg
    Price
    Buy

    Particle Counter/Cell Sizing and Processing - Clinical Diagnostics

    Show: 25 50 100 items per page.

    Info
    Part No.
    Name
    Price
    Buy

    Particle Counter/Cell Sizing and Processing

    Show: 25 50 100 items per page.

    Info
    Part No.
    Name
    Price
    Buy

    Rotors - Clinical Diagnostics

    Show: 25 50 100 items per page.

    Info
    Part No.
    Name
    Max Speed
    Max RCF Atr Maxg
    kFactor
    Price
    Buy

    Rotors

    Show: 25 50 100 items per page.

    Info
    Part No.
    Name
    Max Speed
    Max RCF Atr Maxg
    kFactor
    Price
    Buy

    Tubes & Bottles - Clinical Diagnostics

    Show: 25 50 100 items per page.

    Info
    Part No.
    Name
    Closure Type
    Dimensions
    Material
    Price
    Buy

    Tubes & Bottles

    Show: 25 50 100 items per page.

    Info
    Part No.
    Name
    Closure Type
    Dimensions
    Material
    Price
    Buy
    * All trademarks are property of their respective owners.
    Kaluza Questions & Concerns
    RNACleanXP Certificate of Analysis
    CleanSeq Certificate of Analysis
    AmpureXP Certificate of Analysis
    Quote Request
    Reach Out
    Reach Out Part 2
    Download Resource