Mastitis is the single most costly disease of dairy cattle resulting in the reduction of milk yield and quality. The inflammation of the utter is mainly caused by infection of various bacteria. Streptococcus dysgalactiae, a Lancefield group C α-hemolytic gram-positive bacteria, is a common pathogen in subclinical and clinical mastitis causing substantial economic losses in dairy herds. Streptococcus dysgalactiae has been isolated from infected mammary glands and teat injuries and is transmitted primarily during milking. However, detection of S. dysgalactiae in other cow tissues such as tonsils, mouth and vagina, and occurrence of S. dysgalactiae mastitis during the non-lactating period in herds, suggests that the organism also behaves as an environmental pathogen.
Storage Conditions and Product Stability All kit components can be stored for 2 years after the date of production without showing any reduction in performance.
All kit components should be stored at -20°C upon arrival. Repeated thawing and freezing (> 2 x) of the Master Mix and Positive Control should be avoided, as this may affect the performance of the assay. If the reagents are to be used only intermittently, they should be frozen in aliquots.
Component
Cat. TM30750 (100 preps)
Cat. TM30710 (100 preps)
MDx TaqMan 2X PCR Master Mix
2 x 700 μL
–
S. dysgalactiae Primer & Probe Mix
280 μL
280 μL
S. dysgalactiae Positive Control
150 μL
150 μL
Nuclease-Free Water (Negative Control)
1.25 mL
1.25 mL
Product Insert
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EXTRAClean RNA Clean-Up and Concentration Micro-Elute Kit
Product Info
Document
Product Info
Overview
Ensure optimal results for sensitive applications like NGS
Up to a tenfold increase in microRNA mapping during sequencing runs to reduce costs
Concentration of small amounts of RNA into 8 μL
Ideal for concentrating RNA purified from exosomes, plasma, serum, urine, and other bodily fluids and any RNA samples initially purified in large volumes
Efficient RNA cleanup from enzymatic reactions – labeling, DNase treatment and in vitro transcription
Cleanup of RNA isolated using different methods, including phenol/chloroform extractions
Fast and easy processing using rapid spin-column format
Suitable for all sizes of RNA, from large rRNA down to microRNA (miRNA)
No phenol or chloroform extractions
Purification is based on EXTRAClean spin column chromatography that uses Norgen’s proprietary separation matrix.
Norgen’s EXTRAClean RNA Clean-Up and Concentration Micro-Elution Kit provides a rapid method for the purification, cleanup and concentration of up to 10 μg of RNA isolated using different methods including phenol/guanidine-based protocols, and from various upstream enzymatic reactions such as DNase treatment, labeling and in vitro transcription. The minimum recommended elution volume is 8 μL, which enables the concentration of small amounts of all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA). The RNA is preferentially purified from other reaction components such as proteins, RNases and nucleotides, without the use of phenol or chloroform. The EXTRAClean columns undergo stringent processing and rigorous quality control measures to minimize contamination traces, ensuring optimal results for sensitive applications such as NGS. The purified RNA is of the highest integrity, and can be used in a number of downstream applications including end-point or quantitative reverse transcription PCR, Northern blotting, RNase protection and primer extension, expression array assays and next generation sequencing.
Details
Kit Specifications (Spin Column)
Maximum Column Binding Capacity
10 μg
Size of RNA Purified
All sizes, including miRNA and small RNA (< 200 nt)
All shipping requirements in one convenient mailer
Leak-proof biohazard bags with adsorbent pads for enhanced safety
Matches the standards of OSHA as well as the IATA regulations
Norgen’s Shipping Accessories provides a safe and convenient tool for shipping non-infectious biological specimens. The accessories are contained in individual SHIP TO envelopes that are used for shipping of both the accessories as well as the individual sample collection device to the donor/user.
Each SHIP TO envelope contains:
A Biohazard Specimen Bag with absorbent pad;
A Bubble Envelope labeled as “Exempt Human Specimen”;
2 blank labels;
Shipping Instructions
The Biohazard Specimen Bag is a sure-seal zipper bag to hold the collected specimen and protect against accidental spills. The bag contains an absorbent pad that can adsorb up to 4 mL of liquid and meets current labeling requirements of OSHA standard 29CFR1910.1030 regarding biohazard symbols. The Bubble Envelope is used as a RETURN envelope for the donor/user to ship the collected sample back to the lab for analysis. The user will first place the collected sample into the Biohazard Specimen Bag, and then place the Biohazard Specimen Bag into the Bubble Envelope. The Bubble Envelope protects against moisture and punctures, and facilitates smooth insertion with convenient self-sealing closure. The 2 labels are used for both the outer SHIP TO envelope and the RETURN bubble envelope. The SHIP TO label will be filled out to contain the mailing information of the donor and the RETURN label will be filled out to contain the address to which the sample should be returned. The labels can be supplied as sheets together with the word template file for ease of printing. Norgen’s Shipping Accessories match the regulations of the International Air Transport Association (IATA).
Soil samples contain a large number of microorganisms, the vast majority of which can not be directly cultivated for reproduction and research. Extracting DNA from soil samples is the most effective method for studying soil microorganisms. At present, there are mainly direct and indirect methods for extracting microbial DNA from soil samples. The direct method refers to placing soil samples in the lysis solution, and using effective wall breaking methods to release all microbial DNA into the lysis solution, followed by separation and extraction, such as Zhou’s method. Indirect method refers to placing soil in a buffer, such as Buffer PBS, to separate microorganisms from the soil and then extract DNA. The indirect method can greatly reduce the impact of humic acids and heavy metal salts on DNA extraction in soil, but this method will lose many microorganisms and the resulting DNA is not the entire genome (metagenome) of the soil sample. Currently, few researchers have adopted this method. Extracting DNA directly from soil samples can maximize the likelihood of obtaining the entire genome, but this method faces the following issues:
1. Humic acid pollution. The soil, especially in forests and grasslands, is rich in humic acids. Humic acid is a series of organic molecules, some of which are very similar to nucleic acid molecules and difficult to remove during purification. Trace amounts of humic acid pollution can lead to downstream applications such as PCR and enzyme digestion failure.
2. Lysis method. Soil samples contain various microorganisms, such as bacteria and fungi. Gram positive bacteria and fungi both contain very thick bacterial walls, and effectively breaking down the cell walls of these microorganisms is crucial for extracting high-yield metagenomic DNA. Due to the complexity of soil samples, it is not feasible to use enzymatic methods (such as lysozyme, wall breaking enzyme, snail enzyme) or liquid nitrogen grinding, as the soil contains various metalions or inhibitory factors that inactive the digestive enzymes, or the presence of sand particles in the soil makes liquid nitrogen grinding difficult.
3. The DNA yield is difficult to control. Soil samples would have significant changes in the number and variety of microorganisms due to fertility, inferiority, high moisture content, dryness, or depth of sampling. In a small range of soil samples, the DNA content often varies by thousands of times. In addition, certain chemical components in soil, such as heavy metal salts and clay substances, can cause a decrease in DNA yield.
Magen’s HiPure Soil DNA Kits are currently the most optimized kit for soil DNA extraction. The kit adopts glass bead grinding method and thermal shock chemical wall breaking method, which can be carried out in the point vortex instrument without special bead grinding instrument, and is suitable for a wide range of laboratories. The Absorber Solution in the reagent kit is a humic acid adsorbent exclusively developed by Magen Company, which can efficiently remove various humic acid pollutants. In addition, an alcohol-free silica gel column purification method is also used to efficiently remove various soluble metal salts and other soluble inhibitory factors from the soil. The kit has successfully extracted from the following soil (partially based on customer feedback): soil from forests in nature reserves (30 to 40 years old forest soil with a surface layer of 30-50cm deciduous layer), mangrove soil, grasslands, farmland, seabed mud, sludge, mineral area soil, organic matter contaminated soil, pond mud, garbage mud, air conditioning pipeline deposits, etc.
This product allows rapid and reliable isolation of high-quality genomic DNA from various soil samples. Up to 500 mg soil samples can be processed in 60 minute. The system combines the reversible nucleic acid binding properties of HiPure matrix with the speed and versatilityof spin column technology to eliminate PCR inhibiting compounds such as humic acid from soil samples. Purified DNA is suitable for PCR, restriction digestion, and next-generation sequencing. There are no organic extractions thus reducing plastic waste and hands-on time to allow multiple samples to be processed in parallel.
Details
Specifications
Features
Specifications
Main Functions
Isolation DNA from 200-500mg soil sample
Applications
PCR, southern blot and enzyme digestion, etc.
Purification method
Mini spin column
Purification technology
Silica technology
Process method
Manual (centrifugation or vacuum)
Sample type
Soil
Sample amount
200-500mg
Elution volume
≥30μl
Time per run
≤60 minutes
Liquid carrying volume per column
800μl
Binding yield of column
100μg
Principle
Soil sample is homogenized and then treated in a specially formulated buffer containing detergent to lyse bacteria, yeast, and fungal samples. humic acid,proteins, polysaccharides, and other contaminants are removed using our proprietary Absorber Solution. Binding conditions are then adjusted and the sample is applied to a DNA Mini Column. Two rapid wash steps remove trace contaminants and pure DNA is eluted in low ionic strength buffer. Purified DNA can be directly used in downstream applications without the need for further purification.
Advantages
Fast – several samples can be extracted in 40 minutes (after digestion)
High purity – purified DNA can be directly used in various downstream applications
Good repeatability – silica technology can obtain ideal results every time
High recovery – DNA can be recovered at the level of PG
Kit Contents
Contents
D314202
D314203
Purification Times
50 Preps
250 Preps
Hipure DNA Mini Columns II
50
250
2ml Collection Tubes
50
250
2ml Bead Tubes
50
250
Buffer SOL
60 ml
250 ml
Buffer SDS
5 ml
20 ml
Buffer PS
10 ml
50 ml
Absorber Solution
10 ml
50 ml
Buffer GWP
40 ml
220 ml
Buffer DW1
30 ml
150 ml
Buffer GW2*
20 ml
2 x 50 ml
Buffer AE
15 ml
30 ml
Storage and Stability
Absorber Solution 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.
Experiment Data
Document
Soil samples contain a large number of microorganisms, the vast majority of which can not be directly cultivated for reproduction and research. Extracting DNA from soil samples is the most effective method for studying soil microorganisms. At present, there are mainly direct and indirect methods for extracting microbial DNA from soil samples. The direct method refers to placing soil samples in the lysis solution, and using effective wall breaking methods to release all microbial DNA into the lysis solution, followed by separation and extraction, such as Zhou’s method. Indirect method refers to placing soil in a buffer, such as Buffer PBS, to separate microorganisms from the soil and then extract DNA. The indirect method can greatly reduce the impact of humic acids and heavy metal salts on DNA extraction in soil, but this method will lose many microorganisms and the resulting DNA is not the entire genome (metagenome) of the soil sample. Currently, few researchers have adopted this method. Extracting DNA directly from soil samples can maximize the likelihood of obtaining the entire genome, but this method faces the following issues:
