[RQ2200] ExcelRT™ One-Step RT-qPCR Kit (TaqMan, no ROX), 200 RXN
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The ExcelRT™ One-Step RT-qPCR kit (TaqMan, no ROX) is designed for reverse transcription and quantitative real-time analysis of a specific target RNA by one-step reaction. The ExcelRT™ One-Step RT-qPCR kit (TaqMan, no ROX), consisting of One-Step RT Enzyme Mix and 2X One-Step Master Mix, is a convenient kit designed for highly efficient cDNA synthesis and highly specific real-time PCR in a single tube. The One-Step RT Enzyme Mix contains a thermostable ExcelRT™ Reverse Transcriptase and a RNAok™ RNase inhibitor. Consequently, One-Step RT Enzyme Mix can reverse transcribe RNA to cDNA at a wide temperature range from 42 to 60°C and active against RNase A, RNase B and RNase C. By containing specialized hot-start Taq DNA polymerase, which greatly reduce primer-dimer formation and can be activated within 2 minutes, the 2X One-Step Master Mix features high specificity and is suitable for fast cycle program.
Detail
Description
The ExcelRT™ One-Step RT-qPCR kit (TaqMan, no ROX) is designed for reverse transcription and quantitative real-time analysis of a specific target RNA by one-step reaction. The ExcelRT™ One-Step RT-qPCR kit (TaqMan, no ROX), consisting of One-Step RT Enzyme Mix and 2X One-Step Master Mix, is a convenient kit designed for highly efficient cDNA synthesis and highly specific real-time PCR in a single tube. The One-Step RT Enzyme Mix contains a thermostable ExcelRT™ Reverse Transcriptase and a RNAok™ RNase inhibitor. Consequently, One-Step RT Enzyme Mix can reverse transcribe RNA to cDNA at a wide temperature range from 42 to 60°C and active against RNase A, RNase B and RNase C. By containing specialized hot-start Taq DNA polymerase, which greatly reduce primer-dimer formation and can be activated within 2 minutes, the 2X One-Step Master Mix features high specificity and is suitable for fast cycle program.
Features
Reverse transcription at wide temperature range (42-60°C)
High specificity
Suitable for fast cycle program
With no ROX reference dye
Storage
Aliquot to avoid multiple freeze-thaw cycles (stable within 30 freeze-thaw cycles)
Protect from light
-20°C for 12 months
Other Products
IVD3141 HiPure Stool DNA Kit
Product Info
Document
Product Info
Introduction
With the development of molecular biology, stool, a new non-invasive sample, has been widely used in the research of animal molecular genetics, population ecology, behavioral ecology and some intestinal disease diagnosis. Stool samples includes gut microbial DNA, food residue sample DNA, and alimentary tract exfoliated cell DNA.
The primary problem encountered when using stool sample for molecular biology research is the low content of exfoliated cells in the digestive tract and a certain degree of degradation of genetic material in stool. Another issue in molecular scatology research based on PCR is the presence of a large number of inhibitors in stool that can affect Taq enzyme activity, leading to downstream detection inactivation. These inhibitors include polysaccharides, plant polysaccharides, bile acids, bile salts, bile pigments, digestive juices, mucus, etc. Therefore, selecting appropriate extraction methods to obtain high-quality DNA is the key to successful downstream detection of stool DNA.
At present, the pretreatment methods used in the laboratory, such as phenol/chloroform extraction, cetyltrimethyl bromide (CTAB) lysis, and guanidine isothiocyanate lysis, lack universality in different species, and the success rate of extracting DNA for PCR amplification is also very low. The HiPure Stool DNA Kit provided by Magen Company has opened up a new approach for DNA extraction from stool samples with good universality, high cost-effectiveness, high yield and purification. The reagent kit adopts a unique solution system and inhibitory factor adsorbent, which can efficiently remove various impurities in stool samples. The purified DNA can be directly used for PCR, quantitative PCR and other applications.
This product allows rapid and reliable isolation of high-quality genomic DNA from various stool samples. Up to 100 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 versatility of 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 total DNA from 50-100mg stool samples
Applications
PCR, Southern Blot, enzyme digestion and NGS, etc.
Purification method
Mini spin column
Purification technology
Silica technology
Process method
Manual (centrifugation or vacuum)
Sample type
Stool
Sample amount
50-100mg
Yield
3-15μg
Elution volume
≥30μl
Time per run
≤60 minutes
Liquid carrying volume per column
750μl
Binding yield of column
100μg
Principle
Stool 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
High purity – unique adsorbent can completely remove inhibitory factors
High concentration – maximum extraction of total DNA from stool samples
High recovery – DNA can be recovered at the level of PG
Good repeatability – silica technology can obtain ideal results every time
Kit Contents
Contents
IVD3141
Purification Times
50 Preps
HiPure DNA Mini Columns II
50
2ml Collection Tubes
50
2ml Bead Tubes
50
Proteinase K
24 mg
Protease Dissolve Buffer
1.8 ml
Buffer SPL
40 ml
Buffer PCI
40 ml
Buffer AL
20 ml
Buffer GW1
22 ml
Buffer GW2
20 ml
Buffer AE
15 ml
Storage and Stability
Proteinase K and Buffer PCI 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. 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
With the development of molecular biology, stool, a new non-invasive sample, has been widely used in the research of animal molecular genetics, population ecology, behavioral ecology and some intestinal disease diagnosis. Stool samples includes gut microbial DNA, food residue sample DNA, and alimentary tract exfoliated cell DNA.
Short term stability: 2-8oC, Long term stability: See individual component labels
Stability:
> 2 years under recommended storage conditions
Analyte:
Available Carbohydrates, Dietary Fiber
Assay Format:
Spectrophotometer
Detection Method:
Absorbance
Wavelength (nm):
340
Signal Response:
Increase
Linear Range:
4 to 80 μg of D-glucose, D-fructose or D-galactose per assay
Limit of Detection:
1.475 g/100 g
Reaction Time (min):
~ 5 h
Application examples:
Food ingredients, food products and other materials.
Method recognition:
AOAC Method 2020.07
The Available Carbohydrates Assay Kit method is suitable for the determination of available carbohydrates (AVCHO) comprising *total digestible starch (TDS) plus maltodextrins, sucrose, D-glucose, D-fructose and lactose. New Improved method receiving ‘First Action’ status: AOAC 2020.07. This method is designed to simulate in vivo conditions in the human small intestine (i.e. a 4 h incubation time with PAA + AMG) in parallel with recent advances in Dietary Fiber (DF) methodology (K-RINTDF: AOAC Method 2017.16) and in accordance with the new (physiological based) definition of DF announced by Codex Alimentarius in 2009. Also, sucrose is hydrolysed with a specific “sucrase” enzyme which (unlike invertase which has been used traditionally for this reaction) has no action on fructo-oligosaccharides (FOS).
* Total digestible starch (TDS) is defined as starch that is digested in a 4 h period and is part of the carbohydrate that is available for digestion and absorption in the human small intestine.
The Available Carbohydrates Assay Kit method is suitable for the determination of available carbohydrates (AVCHO) comprising *total digestible starch (TDS) plus maltodextrins, sucrose, D-glucose, D-fructose and lactose. New Improved method receiving ‘First Action’ status: AOAC 2020.07. This method is designed to simulate in vivo conditions in the human small intestine (i.e. a 4 h incubation time with PAA + AMG) in parallel with recent advances in Dietary Fiber (DF) methodology (K-RINTDF: AOAC Method 2017.16) and in accordance with the new (physiological based) definition of DF announced by Codex Alimentarius in 2009. Also, sucrose is hydrolysed with a specific “sucrase” enzyme which (unlike invertase which has been used traditionally for this reaction) has no action on fructo-oligosaccharides (FOS).
SRY (Sex Determining Region Y)-Box 10 (SOX-10), also known as transcription factor SOX-10, is a nuclear transcription factor that acts in regulation of embryonic development and in the specification and differentiation of cells of melanocytic lineage. SOX-10 is diffusely expressed in neurofibromas and schwannomas, and mutations in the SOX-10 gene are linked to Waardenburg-Shah and Waardenburg-Hirschsprung disease. Anti-SOX-10 has been shown to be sensitive for conventional, spindled, and desmoplastic melanoma, and has been used to detect metastatic melanoma and nodal capsular nevus in sentinel lymph nodes.
