CO00020 Midstream Cassette 136x16mm, pink w/lens

Overview

• Robust Lysis Solution processes even the most challenging plant species such as pine needle and grape 
• No phenol extractions
• DNA and all sizes of RNA are recovered, including microRNA
• High quality DNA and RNA are purified simultaneously using the same spin column
• No need to split the lysate​
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

This kit provides for rapid spin column isolation and purification of total RNA and genomic DNA simultaneously from a single plant sample without splitting the lysate. Norgen’s plant lysis solution is highly robust and effective over a wide range of plants including challenging samples.  The total RNA and genomic DNA are both column purified in under 30 minutes.   Since RNA and DNA are isolated without splitting the lysate, variability and inconsistent results are reduced.​​ All sizes of RNA including microRNA are recovered without the need for phenol.  Optional on-column DNase and RNase treatments provide flexibility to isolate DNA-free RNA or RNA-free DNA respectively.  Isolated nucleic acids are of a high quality and yield, and are ready for downstream use including PCR, qPCR, RT-PCR, qRT-PCR, sequencing and more. 

Background

It is often necessary to isolate total RNA and genomic DNA from a single plant sample, such as for studies of gene expression, mutant or transgenic plant characterization, and host plant-pathogen characterization. This is of great benefit when isolating RNA and DNA from precious, difficult to obtain or very small samples. Furthermore, gene expression analysis will be more reliable since the RNA and DNA are derived from the same sample, therefore eliminating inconsistent results.

Details

Supporting Data

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Kit Specifications
Column Binding Capacity	50 μg for RNA 15 μg for genomic DNA
Maximum Column Loading Volume	650 μL
Size of RNA Purified	All sizes, including small RNA (< 200 nt)
Maximum Amount of Starting Material: Plant Tissues Plant Cells	100 mg 5 x 106
Time to Complete 10 Purifications	30 minutes
Average Yields* Peach Leaves (100 mg)	40 μg RNA, 5 μg gDNA

* Yield will vary depending on the type of sample processed

Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. These reagents should remain stable for at least 2 years in their unopened containers.

Component	Cat. 24400 (50 preps)
Lysis Buffer M	40 mL
Binding Buffer I	7 mL
Solution WN	18 mL
Wash Solution A	38 mL
Elution Buffer E	20 mL
Enzyme Incubation Buffer B	6 mL
Filter Columns	50
Spin Columns	50
Collection Tubes	100
Elution Tubes (1.7 mL)	50
Product Insert	1

Introducing the Fastin Assay Kit: Your Straightforward Solution for Elastin Quantification! Our user-friendly kit utilizes a dye-based method to measure elastin from in-vivo and in-vitro sources. It can be used to quantify various elastin forms, spanning from immature tropoelastin to mature, ‘insoluble’ elastin fibers.

Colorimetric Detection (513nm) (Endpoint)

Understanding Elastin: The Key to Tissue Flexibility

Tissues like lungs and arteries must maintain the ability to stretch and recoil repeatedly throughout an organism’s life. Elastin, a mature protein, is responsible for this elasticity and is usually present as insoluble fibers within the ECM. During development, these fibers are initially formed from a soluble precursor called tropoelastin.

‍What is the Fastin Assay?

The Biocolor Fastin assay is a user-friendly, dye-based means of quantifying elastins derived from both in-vivo and in-vitro sources. A variety of elastin forms can be assayed, from immature tropoelastin to mature ‘insoluble’ elastin fibres.

Further information on how the assay works can be found on the ‘Mode of Action‘ tab.

A list of suggested sample types can be found under the ‘Assay Specification‘ tab.

How does the Fastin assay detect Elastin?

The Fastin Dye Reagent contains an elastin-binding synthetic porphyrin, TPPS (5,10,15,20-tetraphenyl- 21H,23H-porphine). This affinity of TPPS for elastin was first observed when used as a ‘vital stain’ on live animals. Most tissues took up the dye initially but only elastin retained the TPPS molecules over time. [Winkelman, J. (1962), Cancer Res. 22, 589-596; Winkelman, J & Spicer, S. (1962), Stain Technol. 37, 303-305].

It has been proposed that the elastin binding of TPPS may be due to the retention of the acidic dye (which contains four charged sulfate groups) by the basic amino acid side chain residues of elastin.

‍How does the Fastin assay work?

Step 1. Incubation of samples containing soluble elastin with the Fastin Dye Reagent causes an elastin-dye complex to form. This insoluble complex then precipiates.

Step 2. Dye-labelled elastin is then isolated by centrifugation and the unbound dye removed. Elastin-bound dye is then eluted and measured spectrophotometrically.

Step 3. The elastin content of unknown samples can be calculated by comparison against a calibration curve prepared using a standard comprising water-soluble elastin (supplied with the kit).

Assay range

50 – 500µg/ml

Limit of Detection

50µg/ml

Detection Method

Colorimetric Detection (513nm) (Endpoint)

Measurements per kit

110 in total (allows a maximum of 48 samples to be run in duplicate alongside a standard curve).

Suitable Samples

In-vivo: tissues and fluids. Insoluble elastin will first require conversion to water soluble α-elastin using the oxalic acid reagents and extraction protocol supplied with the kit.  

In-vitro: Elastin produced by cells during 2D/3D cell culture. NB elastin in conditioned cell media is typically below the detection limit of the kit.

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Precautions

This kit is designed for research use only. Not for use in diagnostic procedures.
Kit requires access to a centrifuge, heated water bath or block, as well as a spectrophotometer or colorimeter capable of absorbance detection at 513nm.
Specific sample preparation protocols may require customer to provide further reagents, consult assay manual for further information.

Fastin elastin kit contents:‍

1. Dye Reagent (1x110ml)

2. α-elastin Reference Standard (1x5ml, 1.0 mg/ml soluble Bovine elastin)

3. Oxalic Acid (1x20ml) Precipitating Reagent (1x30ml)

4. Dye Dissociation Reagent (1x30ml)

5. Precipitating Reagent (1x30ml)

6. 1.5ml micro-centrifuge tubes for dye-labelling reaction.

7. Assay kit manual

NB: Additional reagents may be required for sample preparation prior to assay. Consult manual or contact us for further details.

Introducing the Fastin Assay Kit: Your Straightforward Solution for Elastin Quantification! Our user-friendly kit utilizes a dye-based method to measure elastin from in-vivo and in-vitro sources. It can be used to quantify various elastin forms, spanning from immature tropoelastin to mature, ‘insoluble’ elastin fibers.
Colorimetric Detection (513nm) (Endpoint)