Synovial Fluid Bacterial Genomic DNA Purification Kit
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Detail
Overview
Isolate genomic DNA from all types of bacteria (both Gram-positive and Gram-negative)
Rapid and convenient spin column protocol
Purified bacterial gDNA has a minimal host gDNA contamination.
High yield, high quality DNA for sensitive downstream applications including sequencing, PCR, qPCR and more
Synovial fluid is secreted into the joint cavity from the inner membrane of synovial joints. Synovial fluid is a plasma ultrafiltrate which contains proteins derived from both the blood plasma and produced by cells within the joint tissues. It lubricates the articulating joints as well as supplies oxygen and nutrients while removing carbon dioxide and metabolic wastes from the chondrocytes in the surrounding cartilage. Septic arthritis is usually caused by bacterial, viral or fungal infections to the synovial fluid. Staphylococcus aureus is the most common bacterial infection causing Septic arthritis. Diagnosing Septic arthritis is done through joint fluid (synovial fluid) analysis, blood tests or imaging tests.
Norgen’s Synovial Fluid Bacterial Genomic DNA Isolation Kit provides a fast, reliable and simple procedure for isolating the highest quality and the highest quantity of bacterial genomic DNA from various amounts of synovial fluid ranging from 1 mL up to 10 mL. Purification is based on using Norgen’s proprietary resin separation matrix. The kit is designed to isolate all Gram +ve and Gram -ve strains. Moreover, the kit allows the user to elute the purified bacterial genomic DNA into a flexible elution volume ranging from 25 µL to 50 µL. The purified bacterial gDNA is eluted in an Elution Buffer that is compatible with all downstream applications including PCR, qPCR, methylation-sensitive PCR, Southern Blot analysis and NGS.
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 1 year in their unopened containers.
Component
Cat. 67900 (50 preps)
Precipitation Solution B
15 mL
Lysis Buffer T
6.5 mL
Solution BX
28 mL
Wash Solution A
38 mL
Elution Buffer B
8 mL
Micro Spin Columns
50
Collection Tubes
50
Elution Tubes (1.7 mL)
50
Product Insert
1
Other Products
Sircol-2.0 Soluble 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 SOLUBLE forms of collagen. Chose the Sircol Insoluble collagen kit if you need to analyse INSOLUBLE 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 Sircol 2.0 detect collagen?
The Sircol 2.0 dye reagent includes Sirius Red, a linear anionic dye with sulfonic acid side chains. This reagent is specially formulated to bind to the Gly-X-Yn helical structure of soluble collagen under assay conditions.
*The improved formulation of Sircol 2.0 dye enables a greater degree of dye-collagen specificity (compared to our previous S1000 assay kit).
Overview of the Sircol 2.0 assay process:
Step 1. Prepared samples are placed in the wells of the assay microplate, together with Sircol Dye Reagent. After 30 minutes mixing, any collagen-dye complexes will form as a precipitate. These are collected on the base of the microplate wells by centrifugation.
Step 2. Unbound dye is removed by gentle aspiration, followed by a rinse with Plate Wash Reagent.
Step 3. Following further centrifugation, collagen-bound dye is eluted by incubation with a Dye Release Reagent. Eluted dye is detected ‘in-situ’ by spectrophotometric analysis of the microplate at 556nm.
Step 4. The collagen content of unknown samples can be quantified by comparison against a calibration curve, prepared using the Collagen Reference Standard supplied with the kit.
A list of suggested sample types can be found under the ‘Assay Specification‘ tab.
Colorimetric Detection (556nm) (Endpoint), Requires a microplate centrifuge.
Measurements per kit
96 in total (allows a maximum of 41 samples to be run in duplicate alongside a standard curve).
Suitable Samples
Soluble* collagens from mammalian**:
In-vivo: Tissues, cartilages and fluids.
In-vitro: Extracellular matrices / Conditioned media from 2D/3D culture environments.
The straightforward sample processing and analysis of Sirco 2.0 make it a good alternative to conventional hydroxyproline analysis.
*Prior salt/acid/acid-pepsin extraction may be necessary to release soluble collagen.
**Sircol 2.0 is primarily designed for use with in-vivo / in-vitro samples of mammalian origin. Collagens originating from other taxonomic groups and kingdoms can also be analysed. See note on p6 of manual for further information.
Precautions
This kit is designed for research use only. Not for use in diagnostic procedures. Kit requires access to a microplate centrifuge* (see note below), 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.
*As a minimum, we recommend that the centrifuge can centrifuge a 96-well microplate at 400 x g for 120 minutes. Higher speed centrifuges are recommended (up to a maximum of 2000 x g), allowing a reduction in centrifuge time.
Sircol 2.0 kit contents:
1. Dye Reagent (1x20ml)
2. Collagen Reference Standard (1x5ml, 200µg/ml of soluble Bovine collagen)
3. Plate Wash Reagent (1x28ml)
4. Collagen Concentration Reagent (1x25ml)
5. Neutralisation Reagent (1x8ml)
6. Dye Release Reagent (1x25ml)
7. Assay Microplate (1×96-wells)
8. Microplate Seals (6x)
9. Documentation (QuickStart Guide / Manual / Certificate of Analysis)
NB: Additional reagents may be required for sample preparation prior to assay. Consult manual or contact us for further details. This kit requires the use of a microplate centrifuge, capable of centrifuging a 96-well microplate at 400 x g for 120 minutes. Higher speed centrifuges are recommended (up to a maximum of 2000 x g), allowing a reduction in centrifuge time.
Document
Experience user-friendly detection & measurement of Soluble Collagen with Sircol™ 2.0! Our latest kit simplifies collagen quantification within in-vivo / in-vitro samples. Sircol 2.0 offers enhanced sensitivity and accuracy compared to our previous Sircol kit.
How to use: 1.Suspend 18g in 1L of distilled water, stirring heated to boiling to completely dissolve, autoclave at 121℃ for 15 minutes. 2.Diluted and treated samples.
Quality control:
Item
The name and number of strain
Growth
Colony Color
1
Escherichia coli ATCC25922
Good
Turbid broth,slight precipitate
2
Staphylococcus aureus ATCC25922
Good
Turbid broth,slight precipitate
Storage: Keep container tightly closed, store in a cool, dry place, away from bright light. Storage period of 3 years.
Short term stability: 2-8oC, Long term stability: See individual component labels
Stability:
> 2 years under recommended storage conditions
Analyte:
α-Amylase
Assay Format:
Spectrophotometer, Auto-analyser
Detection Method:
Absorbance
Wavelength (nm):
400
Signal Response:
Increase
Limit of Detection:
0.003 U/g
Reproducibility (%):
~ 3%
Total Assay Time:
~ 30 min
Application examples:
Cereal flours, fermentation broths and other materials.
Method recognition:
AACC Method 22-02.01, AOAC Method 2002.01, ICC Standard No. 303, RACI Standard Method and CCFRA (Flour Testing Working Group Method 0018)
The Ceralpha Method: α-Amylase test kit is suitable for the specific measurement and analysis of α-amylase in cereal grains and fermentation broths (fungal and bacterial).
All reagents stable for > 2 years after preparation
Very specific
Simple format
Mega-Calc™ software tool is available from our website for hassle-free raw data processing
Standard included
Validation of Methods
Document
The Ceralpha Method: α-Amylase test kit is suitable for the specific measurement and analysis of α-amylase in cereal grains and fermentation broths (fungal and bacterial).