The ExcelRT™ One-Step RT-PCR Kit is designed for the reverse transcription and PCR amplification of a specific target RNA from either total RNA or mRNA. The ExcelRT™ One-Step RT-PCR Kit provides the user an alternative to the lengthy two step processes (first strand generation and amplification) by using a single mixture, single tube, one step reaction. The ExcelRT™ One-Step RT-PCR Kit contains a 2X reaction premix consisting of an optimized buffer, dNTPs, Mg2+ and enzyme stabilizer, and a blend of recombinant reverse transcriptase and Taq DNA polymerase. The ExcelRT™ One-Step RT-PCR Kit allows the user to complete the RT-PCR process using a thermocycler in a single reaction setting. The ExcelRT™ One-Step RT-PCR Kit is capable of detecting even trace amounts of target RNA and ideal for target RNA amplification and analysis.
Detail
Description
The ExcelRT™ One-Step RT-PCR Kit is designed for the reverse transcription and PCR amplification of a specific target RNA from either total RNA or mRNA. The ExcelRT™ One-Step RT-PCR Kit provides the user an alternative to the lengthy two step processes (first strand generation and amplification) by using a single mixture, single tube, one step reaction. The ExcelRT™ One-Step RT-PCR Kit contains a 2X reaction premix consisting of an optimized buffer, dNTPs, Mg2+ and enzyme stabilizer, and a blend of recombinant reverse transcriptase and Taq DNA polymerase. The ExcelRT™ One-Step RT-PCR Kit allows the user to complete the RT-PCR process using a thermocycler in a single reaction setting. The ExcelRT™ One-Step RT-PCR Kit is capable of detecting even trace amounts of target RNA and ideal for target RNA amplification and analysis.
The Kit integrates phenol/guanidine-based lysis and silica membrane purification of total RNA. MagZol Reagent, included in the kits, is a monophasic solution of phenol and guanidine thiocyanate, designed to facilitate lysis of samples and inhibit RNases. The high lysis efficiency of the reagent and the subsequent removal of contaminants by organic phase extraction enable extracting up to 200μg total purified RNA from less than 100mg animal, plant, fungal samples,bacteria, cells and other samples.
Details
Specifications
Features
Specifications
Main Functions
Isolation total RNA (include miRNA) from 100mg lipid tissue, tissue, cell, plant, body fluids using columns and MagZol reagent
Applications
RT-PCR, qRT-PCR, Northern hybridization, second generation sequencing
Tissue samples (10-100mg) are homogenized in MagZol Reagent. After addition of chloroform, the homogenate is separated into aqueous and organic phases by centrifugation. The upper, aqueous phase is extracted, and ethanol is added to provide appropriate binding conditions. The sample is then applied to the spin column, where the total RNA (up to 100µg) binds to the membrane and phenol and other contaminants are efficiently washed away. High-quality RNA is then eluted in 30-100µl of RNase-free water.
Advantages
Efficient DNA removal – one step RNA extraction can effectively remove genomic DNA
High quality – one step RNA extraction reagent combined with silica gel column can obtain the highest concentration
Fast – several samples can be extracted in 30 minutes
High applicability – samples including animals, plants, bacteria, cells, etc.
High output – enable to process large samples and the yield can be up to 200μg
Kit Contents
Contents
R413002
D413003
Purification Times
50 Preps
250 Preps
HiPure RNA Mini Columns
50
250
2ml Collection Tubes
50
250
MagZol Reagent
60 ml
270 ml
Buffer RW1
50 ml
200 ml
Buffer RW2*
20 ml
50 ml
RNase Free Water
10 ml
30 ml
Storage and Stability
MagZol Reagent 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 its 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.
Experiment Data
Document
The Kit integrates phenol/guanidine-based lysis and silica membrane purification of total RNA. MagZol Reagent, included in the kits, is a monophasic solution of phenol and guanidine thiocyanate, designed to facilitate lysis of samples and inhibit RNases. The high lysis efficiency of the reagent and the subsequent removal of contaminants by organic phase extraction enable extracting up to 200μg total purified RNA from less than 100mg animal, plant, fungal samples,bacteria, cells and other samples.
It is recommended to use the Isothermal fluorescence detector developed by Amp-future, which is also suitable for fluorescence quantitative PCR apparatus with market known brands.
Kit Storage and term of Validity
Storage term: stored at ≤-20℃,keep away from light, avoid heavy weight and repeated freezing and thawing.
Term of Validity: 14 months
Isothermal nucleic acid Principle Summary
This kit is based on a rapid nucleic acid amplification technology at room temperature and constant temperature: at room temperature and constant temperature (generally 39℃~42℃), with the help of auxiliary proteins and single-strand binding proteins,the recombinase and primers form a complex;Source search and combine the target homology domain, at this time,a D-loop region is formed at the homology position and strand exchange begins;along with the dissociation of the recombinase from the complex,the polymerase also binds to the 3′ end of the primer and begins chain extension. At the same time, relying on the function of exonuclease, adding specific molecular probes designed according to the template, and using fluorescence monitoring equipment can realize real-time monitoring of the amplification process of the target fragment.
Isothermal nucleic acid Product Features
1/ High sensitivity and specificity, short reaction time.
2/ The reagent form is freeze-dried, stable and easy to operate.
Technical Parameters:
Parameters
Details
Product Name
DNA Isothermal Amplification Kit EXO
Manufacturer
Amp-future
Storage Temperature
-20°C
Kit Components
Enzymes, Buffers ,Reagents
Packaging
48 Tests/box
Detection Limit
500-1000copies/µL
Shipping
ICE
Test Time
5-20mins
Isothermal nucleic acid Applications
Suitable for DNA isothermal rapid amplification kit(fluorescent type)
Primer: Require pair of nucleotide primers with the length of 25-35 bp.
Fluorescent Probe:Require the suitable length is 46-52nt.
DNA fluorescent kit reaction temperature is 39 to 42℃ and time is 5-20 minutes.
Notes
1/ Please avoid nucleic acid contamination and set blank control during reaction due to the high sensitivity of the kit.
2/ Please take out the required quantity of MIRA reaction units for the experiment, and put the rest under storage conditions when performing the experiment.
