Convenient optimized on-column DNase treatment using Norgen’s RNA Purification Kits
Also includes protocol for digestion in-solution followed by RNA Clean-Up
Guaranteed RNase-Free
Includes Enzyme Incubation Buffer
Cat. 25710 contains one vial of 1,600 units and Cat. 25720 contains 4 vials (1,600 units/vial)
Norgen’s RNA purification kits isolate total RNA with minimal amounts of genomic DNA contamination. However, for some sensitive downstream applications, it may be desirable to remove all traces of residual DNA. Norgen’s RNase-free DNAse I Kit, with Enzyme Incubation Buffer, can be used for optional on-column DNase digestion with any of Norgen’s RNA purification kits. Alternatively, after isolating total RNA using one of Norgen’s RNA purification kits, the RNA elution can be treated with this DNase I. The RNA can then be purified from the DNase using Norgen’s RNA Clean-Up and Concentration Kit (Cat# 23600), and the RNA can then be used in downstream applications.
Details
Each RNase-Free DNase I Kit is supplied complete with sufficient enzyme and enzyme incubation buffer for 50 or 200 reactions.
Storage Conditions The DNase I provided is in lyophilized form. It is stable for at least 3 months if stored at room temperature. However, it is recommended to store the DNase I vial at 2 – 8ºC (or below) upon receipt to maintain stability beyond 3 months. Buffer DR and Enzyme Incubation Buffer can be stored at room temperature. After reconstitution with Buffer DR (see product manual), the DNase I should be stored at -20ºC. All reagents should remain stable for at least 1 year in their unopened containers at the appropriate storage temperature.
Component
Cat. 25710 (50 rxns)
Cat. 25720 (200 rxns)
DNase I
1 vial / 1,600 units
4 vials (1,600 units/vial)
Buffer DR
1 mL
4 x 1 mL
Enzyme Incubation Buffer
6 mL
4 x 6 mL
Product Insert
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Other Products
Sircol™ – Insoluble Collagen assay kit
Product Info
Document
Product Info
What is Collagen?
Collagen is a fundamental component of the extracellular matrix, and the predominant protein in animals, constituting around 30% of total protein mass. A glycoprotein, it is well known for its triple helical structure. This is formed from three polypeptide α-chains with Gly-X-Y repeating residues (Gly for Glycine, X for proline, and Y for hydroxyproline).
Types of Collagen
Over 28 types of collagens have been identified, with Type I collagen being the most abundant. It’s prevalent in ligaments, tendons, skin, and bone tissue. Its mature, insoluble form grants it remarkable strength, making it vital for the mobility of organisms. Collagen also has biochemical functions, influencing cell growth, proliferation, and differentiation.
This version of the kit is designed to detect and measure INSOLUBLE forms of collagen. Chose our Sircol 2.0 collagen kit if you need to analyse SOLUBLE collagen.
Applications of Collagen
Collagen, with its diverse properties, finds utility in various industries. It plays a role in medicine for wound healing and has an expanding role in tissue engineering and cell culture for biomedical purposes. It’s gaining popularity in the cosmetic industry for skin rejuvenation and is used in chemical formulations and the food industry as a functional food supplement and additive.
How does the Sircol assay detect collagen?
Sircol dye reagent contains Sirius Red – a linear anionic dye with sulphonic acid side chain groups. Under assay conditions the Sircol dye binds the basic groups of soluble collagen molecules. Maximal binding occurs in collagens possessing intact triple helix organisation as the highly ordered Gly-X-Yn helical structure of tropocollagen further contributes to dye binding. This results in a high degree of dye-collagen specificity. Affinity is progressively reduced during heat denaturation 4ºC due to the unwinding of the triple helix and formation of random chains.
Overview of the Sircol assay process:
Step 1. Samples being assayed for insoluble collagen must first undergo a 2-3 hour pre-treatment with Sircol Fragmentation reagent. This converts insoluble collagen into water-soluble gelatin can then be assayed.
Step 2. Addition of Sircol Dye Reagent to these pre-treated insoluble collagen samples results in the formation of a denatured collagen-dye complex. This complex then precipitates during the dye incubation period and is subsequently isolated by centrifugation, followed by washing to remove unbound dye. The Denatured collagen-bound dye is then eluted and measured spectrophotometrically.
Step 3. The insoluble collagen content of unknown samples is quantified by comparison against a calibration curve prepared using a the denatured collagen standard supplied with the kit.
Assay range
100 – 1000 µg/ml
Limit of Detection
100µg/ml
Detection Method
Colorimetric Detection (556nm) (Endpoint)
Measurements per kit
110 in total (allows a maximum of 46 samples to be run in duplicate alongside a standard curve).
Suitable Samples
The assay can be used to assess the rate of production of newly laid down collagen fibres during periods of rapid growth, development, tissue repair, remodeling and wound healing. Sources of material includes tissues, bone and calcified tissue.
*Insoluble collagens must be converted into soluble form prior to assay. Instructions and regents are provided with the kit., depending on sample this will require prior salt/acid/acid-pepsin extraction.
**non-mammalian collagens may result in a reduced limit of detection. We recommend use of an assay standard matched to the species under assay.
Many customers have found that the straightforward sample processing and analysis of Sircol make it a good alternative to conventional hydroxyproline analysis.
Precautions
This kit is designed for research use only. Not for use in diagnostic procedures. Kit requires access to a centrifuge, water bath / heated block, as well as a spectrophotometer/colorimeter capable of absorbance detection at 556nm. Specific sample preparation protocols may require customer to provide further reagents, consult assay manual for further information.
Sircol Insoluble Collagen kit contents:
1. Sircol Dye Reagent (1x110ml)
2. Denatured Collagen Reference Standard (1x5ml, 1.0mg/ml)
3. Acid-Salt Wash Reagent (1x20ml)
4. Fragmentation Reagent (1x110ml)
5. Alkali Reagent (1x110ml)
6. 2ml screw-cap tubes for preparation of samples.
7. Assay kit manual
NB: Additional reagents may be required for sample preparation prior to assay. Consult manual or contact us for further details.
Document
As collagens mature, they become increasingly crosslinked and insoluble – characteristics necessary for key biophysical role that collagen plays in living organisms. Biocolor’s Sircol™ INSOLUBLE Collagen Kit is a dye-binding assay designed for accurate quantification and measurement such collagens. It is ideal for analyzing crosslinked / insoluble collagens from sources such as tissues, bone, and calcified tissue.
This product provides high quality purification of total DNA from whole blood, plasma, serum, buffy coat, or other body fluids, lymphocytes and cultured cells. There is no need to use toxic phenol chloroform extraction or time-consuming alcohol precipitation. The extraction process finish in 60 minutes. Purified DNA includes genomic DNA, mitochondrial DNA, viral DNA (e.g. HBV), or DNA from other parasitic microorganisms. The obtained DNA can be directly used in PCR, viral DNA detection and other experiments.
This kit can use on manual protocol or 96 channel automated extraction system.
Details
Specifications
Features
Specifications
Main Functions
Isolation total DNA from 200μl whole blood
Applications
PCR, southern bolt and virus detection, etc
Purification technology
Magnetic beads technology
Process method
Manual or automatic
Sample type
Anticoagulant blood, concentrated blood, buffy coat, lymphocytes and cultured cells
Sample amount
200μl
Elution volume
≥50μl
Time per run
≤60 minutes
Principle
This product is based on the purification method of high binding magnetic particles. The sample is lysed and digested under the action of lysate and Protease. DNA is released into the lysate. After adding magnetic particles and binding solution, DNA will be adsorbed on the surface of magnetic particles, and impurities such as proteins will be removed without adsorption. The adsorbed particles were washed with washing solution to remove proteins and impurities, washed with ethanol to remove salts, and finally DNA was eluted by Elution Buffer.
Advantages
High binding force – suitable for handling DNA rich samples, such as whole blood, buffy coat, concentrated blood, etc.
Fast – polydisperse magnetic beads, fast magnetic response and short extraction time
High purity – the obtained DNA can be directly used for second-generation sequencing, PCR based detection, gene bank, etc.
Automatic – saving time and labor and safer
Kit Contents
Contents
D631101
D631102
D631103
Purification Times
48
96
480
MagPure Particles
1.2 ml
2.5 ml
11 ml
Proteinase K
24 mg
50 mg
220 mg
Protease Dissolve Buffer
1.8 ml
5 ml
15 ml
Buffer AL
15 ml
30 ml
120 ml
Buffer GW1*
22 ml
53 ml
220 ml
Elution Buffer
15 ml
30 ml
100 ml
Storage and Stability
Proteinase K, MagPure Particles should be stored at 2-8°C upon arrival. However, short-term storage (up to 24 weeks) at room temperature (15-25°C) does not affect their performance. The remaining kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. The entire kit can be stored at 2-8°C, but in this case buffers should be redissolved before use. Make sure that all buffers are at room temperature when used.
Document
This product provides high quality purification of total DNA from whole blood, plasma, serum, buffy coat, or other body fluids, lymphocytes and cultured cells. There is no need to use toxic phenol chloroform extraction or time-consuming alcohol precipitation. The extraction process finish in 60 minutes. Purified DNA includes genomic DNA, mitochondrial DNA, viral DNA (e.g. HBV), or DNA from other parasitic microorganisms. The obtained DNA can be directly used in PCR, viral DNA detection and other experiments.
This kit can use on manual protocol or 96 channel automated extraction system.
Attogene Fluorescent UniversalLateral Flow Assay Kits are a convenient ready-to-use kit for quick and cost-effective development of a fluorescent lateral flow dipstick assay for detection of DNA and RNA products.
Fluorescent lateral flow assays can be 2-100 fold more sensitive that gold based lateral flow tests. Europium dyed polystyrene beads conjugated to streptavidin
Formats (fluorescent broad range UV light excitation range of 300nm to 400nm, 610nm emission) Streptavidin conjugate pad):
• Detectable using a black light such as a black light UV flashlight or fluorescent lateral flow reader.
• Detection of nucleic Acid (DNA or RNA) requires the use of a biotin and FAM-labelled primer during amplification.
• Test line: anti-FITC/FAM
• Control Line: Biotin
• Multiplex detection of nucleic Acid (DNA or RNA) requires the use of a biotin, FAM and Dig labelled primers during amplification.:
• Test Line #1: anti FITC/FAM, Line #2: anti-Dig, Line #3 Biotin.
Inquire about custom configurations: sales@attogene.com.
For example, this product can be configured with alternative/custom streptavidin fluorescent particles.
Kit Components
50 -4.5mm Fluorescent Lateral Flow Dipsticks
10 mL Sample assay running buffer
Features & Benefits
Can be used for development of a lateral flow assay for detection of a variety of different molecules such as amplified DNA products from PCR, LAMP and RPA reactions.
No need to stripe capture antibodies
No expensive equipment required (Black Light)
Cost-effective way to screen for further downstream lateral flow assay development.
• Detectable using a black light such as a black light UV flashlight or fluorescent lateral flow reader.