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
NEST 1.5 mL Microcentrifuge Tube (500 count)
Product Info
Document
Product Info
NEST 1.5 mL Microcentrifuge Tube, polypropylene, sterile, DNase, RNase, pyrogen and endotoxin free. Relative centrifugal force (RCF): 30000 x g. Easy-open, secure safelock cap seal. 500 count.
Labware definition is available for immediate use in Opentron’s Labware Library.
Document
NEST 1.5 mL Microcentrifuge Tube, polypropylene, sterile, DNase, RNase, pyrogen and endotoxin free. Relative centrifugal force (RCF): 30000 x g. Easy-open, secure safelock cap seal. 500 count.
Labware definition is available for immediate use in Opentron’s Labware Library.
Blood samples contain rich DNA, including mitochondrial DNA, genomic DNA, circulating DNA (mostly released into blood after tumor cell apoptosis) in white blood cells, as well as parasitic viral or microbial DNA. These DNA are important parameters in clinical testing or diagnosis, which are also valuable materials for medical research. There are three main issues with extracting DNA from blood samples:
1. The sample is highly infectious, posing great harm to operators and the environment.
2. The source of DNA is complex and aportion of the nucleic acid, such as viral DNA or free DNA, may be lost during the operation, leading to downstream detection failure;
3. Blood sample contains a large amount of impurities and inhibitory factors.
Currently there are many methods available for extracting DNA from whole blood samples, such as phenol chloroform extraction, salting out method, etc. However, these methods require pre-treatment of blood sample, which removes red blood cells and isolate white blood cells in the first step. Due to the requirement that it cannot inactivate or kill pathogens during the process of removing red blood cells, the waste liquid (red blood cell lysate) and consumables may be contaminated by pathogens and become infectious, posing a danger to the entire laboratory environment and operators. In addition, during the process of removing red blood cells, useful nucleic acid information such as viruses, microorganisms, or circulating DNA is also lost, leading to experiment or detection failures.
The HiPure Blood DNA Kits series provided by Magen Company uses silica gel column purification technology, which can directly lyse whole blood samples without the need for white blood cell separation. Whole blood samples are directly mixed with lysates and proteases, resulting in the inactivation of pathogens, greatly reducing the infectivity, environmental pollution, and the chance of operators being infected. Due to the direct lysis and digestion of samples, except lymphocyte DNA, other circulating DNA as well as DNA from viruses and microorganisms, can also be recovered.
This product provides fast and easy methods for purification of total DNA for reliable PCR and Southern blotting. Total DNA(e.g., genomic, viral, mitochondrial) can be purified from whole blood (fresh or frozen), plasma, serum, buffy coat, bone marrow, other body fluids, lymphocytes, cultured cells.
Details
Specifications
Features
Specifications
Main Functions
Isolation total DNA from 200ul Whole Blood
Applications
PCR, southern bolt and virus detection, etc
Purification method
Mini spin column
Purification technology
Silica technology
Process method
Manual (centrifugation or vacuum)
Sample type
Whole Blood (fresh or frozen), serum, plasma, milk, saliva, and other liquid samples and cultured cells
Sample amount
<200μl whole blood or other liquid samples, <5*106 lymphocytes or Culture CellsNon mammalian animals that have nucleus in red blood cells (rich in DNA, such as birds and fish) : 5~20μl whole blood at a time
Elution volume
≥20μl
Time per run
≤30 minutes
Liquid carrying volume per column
800µl
Binding yield of column
100µg
Principle
This product is based on silica Column purification. The sample is lysed and digested with lysate and protease, DNA is released into the lysate. Transfer to an adsorption column. Nucleic acid is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, Nucleic acid was finally eluted with low-salt buffer (10mm Tris, pH9.0, 0.5mm EDTA).
Advantages
High quality DNA – meet a variety of downstream applications, including PCR, qPCR, enzyme digestion, hybridization, etc.
Fast – without separation of leukocytes, organic extraction or ethanol precipitation
Simple – all nucleic acids can be obtained by direct digestion
Wide applicability- handle a variety of liquid samples
Kit Contents
Contents
D311102
D311103
Purification Times
50
250
HiPure DNA Mini Columns I
50
2 x 125
2ml Collection Tubes
100
5 x 100
Buffer AL
15 ml
60 ml
Buffer DW1
30 ml
150 ml
Buffer GW2*
12 ml
50 ml
Proteinase K
24 mg
120 mg
Protease Dissolve Buffer
1.8 ml
10 ml
Buffer AE
15 ml
60 ml
Storage and Stability
Proteinase K should be stored at 2–8°C upon arrival. However, short-term storage (up to 12 weeks) at room temperature (15–25°C) does not affect their performance. The remaining kit components can be stored at room temperature (15–25°C) and are stable for at least 18 months under these conditions.
Blood samples contain rich DNA, including mitochondrial DNA, genomic DNA, circulating DNA (mostly released into blood after tumor cell apoptosis) in white blood cells, as well as parasitic viral or microbial DNA. These DNA are important parameters in clinical testing or diagnosis, which are also valuable materials for medical research. There are three main issues with extracting DNA from blood samples: