FP313-FastKing RT-qPCR Buffer (SYBR Green)

FastKing One Step RT-qPCR Kit (SYBR Green)
For real-time RT-qPCR using SYBR Green

FastKing One Step RT-qPCR Kit
(SYBR Green)
Cat. no. 4993103

Storage Conditions:
This kit should be stored at -30~-15°C for one year and protected from
light.
Compatible Real-Time Instruments
ABI PRISM 7000/7700/7900HT, 7300/7500, 7500 Fast, ViiA 7,
QuantStudioTM, StepOneTM/ StepOne PlusTM, 12K Flex (Applied
Biosystems)
OPTICONTM, CFX series (BIORAD)
Smart Cycler® System (Cepheid)
Mx3000 P/Mx3005P (Stratagene)
Line-Gene (Bioer)
Roche series
And others

Introduction
FastKing One Step RT-qPCR Kit is designed for Real-Time One Step RT-qPCR
by using SYBR Green I. The kit can be used in real-time one step RT-qPCR of
RNA targets, with reverse transcription and PCR taking place sequentially in
the same tube. Therefore the operation is simple, and it minimizes the risk
of contamination.
The 25× RT-PCR Enzyme Mix in this kit is a premix of TIANGEN’s novel
reverse transcriptase (FastKing RTase), novel antibody modified hot start
TaqDNA polymerase and RNase Inhibitor. The FastKing RTase is a new
type of molecular modified reverse transcriptase. With the specifically
added hydrophobic motif, which has stronger RNA affinity and thermal
stability. The FastKing RTase has improved reverse transcription efficiency
and the ability to extend the RNA template with complex secondary
structure. The new type of hot start TaqDNA polymerase with excellent
performance is used in the PCR process, making the PCR reaction after
reverse transcription have higher amplification efficiency and specificity. In
addition, the 2× FastKing RT-qPCR Buffer (SYBR Green) in this product is a
new reaction system specially optimized for the above two key enzymes,
including necessary ionic components, dNTPs, SYBR Green dye, one step
stabilizer and enhancer, which can ensure that FastKing RTase and the new
hot start TaqDNA polymerase can play the most effective role in the whole
one-step reaction process.
This kit provides good standard curves in a wide range of quantitative
area. It can accurately and quantitatively detect a variety of high and low
abundance target genes with good repeatability and high reliability.

Product Feature

  1. Increased reaction efficiency: The high-quality reverse transcriptase and
    Taqpolymerse ensure the high reaction efficiency.
  2. Easy and fast operation: The dual component form makes the procedure
    easy and fast.
  3. Capable of reading through complex templates: Suitable for RNA
    templates with high GC content and complex secondary structure.
  4. High compatibility: Compatible for RNA templates from different sample
    source or with impurities.

Reagents and Materials not Supplied

  1. Specific primers
  2. Template RNA
  3. Disposal gloves and other consumables

Application
The RT-qPCR can be used to detect gene expression and RNA virus content
in cells and tissues.
Protocol

  1. Fully thaw the template RNA, specific primers, 2× FastKing RT-qPCR
    Buffer (SYBR Green), 50× ROX Reference Dye and RNase-Free ddH2O,
    and place on ice after short-spinning.
  2. Prepare a reaction mix on ice according to the following table.
  1. Run the Real Time One Step RT-qPCR reaction
    Centrifuge briefly before starting real-time PCR. It is recommended to
    run the standard PCR reaction program in the following table. If the
    program can’t generate optimal results, it is recommended to perform
    further optimization.
  1. Result analysis
    After the reaction, confirm the amplification curve, melting curve,
    CT value and standard curve of the Real Time One Step RT-qPCR, and
    analyze the RT-qPCR quantitative results.

Important Notes

  1. Total RNA or mRNA can be used as RNA template. It is recommended
    to use TRNzol, RNAprep Pure or RNA Easy Fast series produced by
    TIANGEN company to prepare high quality total RNA.
  2. RNase contamination should be avoided during the one-step RT qPCR
    experiment:
    1) Since RNase is found on the surface of human skin and saliva,
    disposable gloves and masks should be worn in the experiment;
    2) Special instruments and consumables should be used in the one
    step RT qPCR experiment, and it is suggested that RNA should be
    operated in special areas;
    3) One step RT-qPCR experiment related consumables should be
    treated with 0.1% DEPC (diethyl pyrocarbonate) solution at 37°C for
    12 hours and autoclaved for 30 min.
  3. The 25× RT-PCR Enzyme Mix should be centrifuged for a short time
    before pipetting. The action should be slow, and it should be put back
    to -30~-15°C as soon as possible after use.
  4. 2× FastKing RT-qPCR Buffer (SYBR Green) should be fully mixed and
    centrifuged before use.

FP313-FastKing RT-qPCR Buffer (SYBR Green)

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This product is for scientific research use only. Do not use in medicine, clinical treatment, food or cosmetics.

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