NEC express5800 A2010b User Manual
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Summary of Contents for NEC express5800 A2010b
- Page 1 RR036Q Cat. # For Research Use PrimeScript™ RT Master Mix (Perfect Real Time) Sample Product Manual v201204Da_2...
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Page 2: Table Of Contents
Cat. #RR036Q PrimeScript™ RT Master Mix （Perfect Real Time） v201204Da_2 Table of Contents Description ......................3 Components ....................... 3 III. Materials Required but not Provided ............4 IV. Storage ......................... 4 Features ........................ 4 VI. Precautions for Use ..................4 VII. -
Page 3: Description
Cat. #RR036Q PrimeScript™ RT Master Mix （Perfect Real Time） v201204Da_2 Description This product is a reverse transcription reagent kit designed to perform reverse transcription optimized for 2 step real-time RT-PCR (RT-qPCR). It contains a 5X pre- mixed reagent containing all of the components needed for quantitative RT-PCR reverse transcription (PrimeScript RTase, RNase Inhibitor, Random 6mers, Oligo dT Primer, dNTP Mixture, and reaction buffer), and a reaction can be started simply by adding template RNA and water. -
Page 4: Materials Required But Not Provided
Cat. #RR036Q PrimeScript™ RT Master Mix （Perfect Real Time） v201204Da_2 III. Materials Required but not Provided - Thermal cycler (or 37℃ water bath and 85℃ heat block) - 0.2 ml and 1.5 ml microtubes (for reverse-transcription reaction) - Micropipettes and tips (autoclaved) IV. -
Page 5: Protocol
Cat. #RR036Q PrimeScript™ RT Master Mix （Perfect Real Time） v201204Da_2 VII. Protocol Reverse transcription reaction For RNA preparation method, please see Section X. 1. Prepare the reverse-transcription reaction solution on ice. <Per reaction> Reagent Amount Final conc. 5X PrimeScript RT Master Mix (Perfect Real Time) 2 μl ＊... -
Page 6: Real-Time Pcr
Cat. #RR036Q PrimeScript™ RT Master Mix （Perfect Real Time） v201204Da_2 VIII. Real-Time PCR The protocol for performing real-time PCR using SYBR® Premix Ex Taq II (Tli RNaseH Plus) (Cat. #RR820A) after performing a reverse-transcription reaction with this kit is shown below. If real-time PCR is to be performed with TaqMan®... - Page 7 Cat. #RR036Q PrimeScript™ RT Master Mix （Perfect Real Time） v201204Da_2 2. Initiate the reaction. The recommended protocol for PCR is the shuttle PCR standard protocol described below. Try this protocol first and optimize PCR conditions as necessary. Perform a 3 step PCR when using a primer with low T value or when a shuttle PCR is not feasible.
- Page 8 Cat. #RR036Q PrimeScript™ RT Master Mix （Perfect Real Time） v201204Da_2 < Method Using Thermal Cycler Dice Real Time System II > 1. Prepare the PCR reaction mixture. <Per reaction> Reagent Amount Final conc. SYBR® Premix Ex Taq II (Tli RNaseH Plus) (2X) 12.5 μl ＊...
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Page 9: Experimental Example
Cat. #RR036Q PrimeScript™ RT Master Mix （Perfect Real Time） v201204Da_2 IX. Experimental Example Reverse-transcription reaction time and amount of cDNA synthesized. [ Method ] 1. Reverse-Transcription Reaction Reagent: PrimeScript RT Master Mix (Perfect Real Time) Template: Human Placenta total RNA 2 pg - 2 μg and sterilized water Reaction volume: 20 μl Reaction conditions: 37℃... -
Page 10: Appendix
Cat. #RR036Q PrimeScript™ RT Master Mix （Perfect Real Time） v201204Da_2 Appendix Preparation of RNA Samples It is important to use a highly pure RNA samples for better cDNA yield. It is essential to inhibit RNase activity in the cells and also to prevent RNase derived from equipment and solutions used. - Page 11 Cat. #RR036Q PrimeScript™ RT Master Mix （Perfect Real Time） v201204Da_2 (2) After extraction of total RNA, remove genomic DNA by Recombinant DNase I (RNase-free) (Cat. #2270A) treatment. After the reaction, DNase I should be inactivated by either heat treatment or phenol/chloroform extraction. Procedure 1.
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Page 12: Related Products
Cat. #RR036Q PrimeScript™ RT Master Mix （Perfect Real Time） v201204Da_2 XI. Related Products SYBR® Premix Ex Taq ™ II (Tli RNaseH Plus) (Cat. #RR820A/B) SYBR® Premix Ex Taq ™ (Tli RNaseH Plus) (Cat. #RR420A/B) Premix Ex Taq ™ (Probe qPCR) (Cat. #RR390A) EASY Dilution (for Real Time PCR) (Cat.