The Kit provides fast purification of high-quality RNA from whole blood, cells and tissues using silica-membrane spin columns with a binding capacity of 100μg RNA. There is no need for phenol / chloroform extractions and time-consuming steps such as CsCl gradient ultracentrifugation or isopropanol precipitation. The purified RNA can be used for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection and in vitro translation.
Specifications
Features | Specifications |
Main Functions | Isolation total RNA from 1-1.5ml whole blood |
Applications | qPCR / RT-PCR, liquid or solid-phasechip analysis, hybridization and SNP detection |
Purification method | Mini spin column |
Purification technology | Silica technology, acidphenol / guanidine extraction technology (MagZol pretreatment technology) |
Process method | Manual (centrifugation or vacuum) |
Sample type | Fresh or frozen blood, mammalian blood |
Sample amount | ≤1.5 ml whole blood |
Yield | 2-100μg |
Elution volume | ≥20μl |
Liquid carrying volume per column | 800µl |
Binding yield of column | 100μg |
The Kit simplifies isolation of RNA from blood with a fast spin-column procedure. Red blood cells are selectively lysed and white cells collected by centrifugation. White cells are then lysed using highly denaturing conditions which immediately inactivate RNases. After homogenization using the DNA spin column, the sample is applied to the RNA column. Total RNA binds to the membrane and contaminants are washed away, leaving pure RNA to be eluted in 30–100µl RNase-free water (provided with the kit) for direct use in any downstream application.
Advantages
Kit Contents
Contents | R416102 | R416103 |
Purification Times | 50 Preps | 250 Preps |
HiPure DNA Mini Columns | 50 | 250 |
HiPure RNA Mini Columns I | 50 | 250 |
2ml Collection Tubes | 100 | 500 |
10 x Buffer RBC | 50 ml | 3 x 100 ml |
RTL Lysis Buffer | 50 ml | 250 ml |
Buffer RW1 | 50 ml | 250 ml |
Buffer RW2* | 20 ml | 2 x 50 ml |
RNase Free Water | 10 ml | 30 ml |
Storage and Stability
HiPure Blood RNA Mini Kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. Make sure that all buffers are at room temperature when used. During shipment, crystals or precipitation may form in the RTL Lysis Buffer. Dissolve by warming buffer to 37°C.
The Kit provides fast purification of high-quality RNA from whole blood, cells and tissues using silica-membrane spin columns with a binding capacity of 100μg RNA. There is no need for phenol / chloroform extractions and time-consuming steps such as CsCl gradient ultracentrifugation or isopropanol precipitation. The purified RNA can be used for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection and in vitro translation.
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.
Streptococcus dysgalactiae TaqMan PCR Kit, 100 reactions
Streptococcus dysgalactiae TaqMan PCR Probe/Primer Set and Controls, 100 reactions
For research use only and NOT intended for in vitro diagnostics.
Figure 1 / 3
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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 | 1 | 1 |
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