Norgen’s 2X One-Step RT-PCR Master Mix is a ready-to-use solution that contains components required for RT-PCR amplification of RNA templates. The mix includes M-MuLV reverse transcriptase, Taq DNA polymerase, dNTPs, reaction buffer, MgCl2, KCl, and a PCR enhancer/stabilizer. The user needs only to add the template, the primer set and water to the Master Mix to set up the RT-PCR reaction. This convenient 2X One-Step RT-PCR Master Mix reduces the time required to set up PCR reactions and reduces the possibility of contamination, particularly when preparing large numbers of reactions. The optimized master mix allows for robust amplification of RNA templates with high yields of PCR products.
Taq DNA Polymerase is a highly thermostable DNA polymerase that possesses a 5´→ 3´ polymerase activity and a very low 5´→ 3´ exonuclease activity. The source of Taq included with Norgen’s 2X One-Step RT-PCR Master Mix is an E. coli strain with a cloned Taq DNA Polymerase gene from Thermus aquaticus YT-1. M-MuLV Reverse Transcriptase is an RNA-directed DNA polymerase that can synthesize a complementary DNA strand initiating from a primer using either RNA (cDNA synthesis) or single-stranded DNA as a template. The source of the Reverse Transcriptase included with Norgen’s 2X One-Step RT-PCR Master Mix is an E. coli strain with a cloned reverse transcriptase gene from M-MuLV.
Norgen’s 2X One-Step RT-PCR Master Mix is available in 3 convenient sizes:
Cat # 28113 – 100 reactions (sufficient for 100 reactions x 20 µL reaction volume)
Cat # 28114 – 200 reactions (sufficient for 200 reactions x 20 µL reaction volume)
Cat # 28115 – 500 reactions (sufficient for 500 reactions x 20 µL reaction volume)
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Reagents supplied – Cat # 28114
Storage Conditions and Product Stability
2X One-Step RT-PCR Master Mix should be stored at -20°C. For everyday use an aliquot can be stored at 4°C for up to 3 months. Repeated freeze-thaw cycles are not recommended. When stored at the proper temperature this reagent is stable for at least 1 year.
HiPure FFPE RNA Kit supplies a simple and rapid RNA extraction for Formalin-fixed, paraffin-embedded (FFPE) tissue and sections samples. This kit is based on silica gel column purification technology, no phenol-chloroform extraction or alcohol precipitation. The whole extraction only takes 30 minutes (not including digestion time). RNA can be directly used for downstream applications such as RT-PCR, Northern blot, vitro translation and other experiments.
Specifications
Features | Specifications |
Main Functions | Isolation total RNA from FFPE tissue |
Applications | RT-PCR, cDNA synthesis, second generation sequencing |
Products | RNA, miRNA |
Purification method | Mini spin column |
Purification technology | Silica technology, DNase |
Process method | Manual (centrifugation or vacuum) |
Sample type | FFPE slice, FFPE embedded tissue |
Sample amount | Tissue: <6 mgFFPE: <6 slices |
Yield | 1-20μg |
Elution volume | ≥15μl |
Time per run | ≤30 minutes (after digestion) |
Liquid carrying volume per column | 800μl |
This product is based on silica column purification. Remove paraffin by Buffer DPS. Sample lysis withproteinase K digestion requires only 15 minutes. After lysis, samples are incubated at 80ºC for 15 minutes. Transfer to an adsorption column and RNA is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, RNA wasfinally eluted with low-salt buffer.
Advantages
Kit Contents
Contents | IVD4144 |
Purification Times | 50 Preps |
HiPure RNA Micro Columns | 50 |
2ml Collection Tubes | 50 |
Proteinase K | 24 mg |
Protease Dissolve Buffer | 1.8 ml |
DNase I | 600 μl |
DNase Booster Buffer | 1.5 ml |
Buffer DPS | 60 ml |
Buffer FRL | 15 ml |
Buffer RLC | 15 ml |
Buffer RWC* | 10 ml |
Buffer RW2* | 20 ml |
RNase Free Water | 10 ml |
Storage and Stability
Proteinase K should be stored at 2–8°C upon arrival. DNase I should be stored at -20°C. However, short-term storage (DNase I up to 1 weeks, Proteinase K up to 8 weeks) at room temperature(15–25°C) does not affect their performance. The remaining kit components can be stored at roomtemperature (15–25°C) and are stable for at least 18 months under these conditions.
Experiment Data
HiPure FFPE RNA Kit supplies a simple and rapid RNA extraction for Formalin-fixed, paraffin-embedded (FFPE) tissue and sections samples. This kit is based on silica gel column purification technology, no phenol-chloroform extraction or alcohol precipitation. The whole extraction only takes 30 minutes (not including digestion time). RNA can be directly used for downstream applications such as RT-PCR, Northern blot, vitro translation and other experiments.
Propargyl-PEG4-sulfonic acid is heterobifunction reagent that can reacts with azide compounds or biomolecules via copper catalyzed Click Chemistry reactions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Propargyl-PEG4-sulfonic acid is heterobifunction reagent that can reacts with azide compounds or biomolecules via copper catalyzed Click Chemistry reactions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
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