One-step RT-qPCR
Inquire-
Features
● No cross-contamination: Reverse transcription and fluorescence quantification are completed in one step to avoid cross contamination.
● Sensitive: Templates as low as 1 ng can be accurately identified, especially suitable for templates with low abundance.
● Stress resistance: Can be applied on complex templates, perfect against foreign interference, suitable for various templates.
● Simple operation: Optimize product composition and improve experimental efficiency.
-
Description
This product is a special reagent for one-step RT-qPCR using probe method (TaqMan, Molecular Beacon, etc.). The Real Time One Step RT-qPCR reaction can be continuously carried out in the same reaction tube using the product. The operation is simple. The cross contamination between samples can be avoided and the detection sensitivity is improved. The kit adopts a premix of TIANGEN novel reverse transcriptase (King RTase), novel antibody modified hot start Taq DNA polymerase and RNase Inhibitor, and has stronger RNA affinity, thermal stability, higher amplification efficiency and specificity. In addition, the 2× FastKing One Step Probe RT-qPCR MasterMix in this product is a new type of reaction system specially optimized for the abovetwo key enzymes. It contains necessary ionic components, dNTPs, PCR stabilizers and enhancers, which can ensure the best performance of King RTase and the new type of hot start Taq DNA polymerase in the whole one-step reaction process.
-
Kit Contents
-
Applications
The kit is suitable for probe one-step Real-Time RT-PCR. It can accurately and simply perform mRNA expression analysis. It is especially suitable for detection of trace RNA on various real time PCR instruments.
-
Required Reagents
● Primers and probes
● Templates
● Disposable gloves and other laboratory supplies
-
Storage Condition
FastKing One Step RT-qPCR Kit could be stored at -30~-15°C for 1 year. -
Important Notes
●The RNA template should be total RNA or mRNA. We recommend using TIANGEN TRNzol reagent or RNAprep kit to extract high-quality RNA template.
● To avoid RNase contamination, the operator must: i. Put on disposable gloves and breathing mask. ii. Use RNase-free material such as reagents, pipette tips, microtubes and instruments. iii. Do the experiments in certain area that especially for RNA operation.
● 25× FastKing Enzyme Mix should be centrifuged transiently before use and slowly pipetted. Put it back to -30~-15°C soon after use.
● Completely mix the 2× FastKing One Step Probe RT-qPCR Mix and centrifuge the reagent to the bottom of the tube before use.
● Use specific reverse transcription primer only. Random Primer and Oligo dT Primer can not be used in reverse transcription reaction.
● To perform several Real Time One Step RT-qPCR at the same time, a mixture of all reagents should be firstly prepared and then divided into each reaction tube. It reduces reagent loss, avoids repeatedly adding the same reagents, and reduces the error by adjusting the volume of each components.
-
Experimental Example
Total RNA of human 293T cells, corn leaves and enteroviruses were extracted respectively and the housekeeping genes were reverse transcribed and quantified by FastKing One Step RT-qPCR Kit (Probe) and relevant product from Supplier A respectively in one step. The results show that TIANGEN FastKing One Step RT-qPCR Kit (Probe) has high fluorescence value, lower CT value, clear gradient and good reproducibility, which obviously outperforms the product from Supplier A.
A: 293T cells. Orange line: TIANGEN, Red line: Supplier A.
B: Corn leaf. Blue line: TIANGEN, Green line: Supplier A.
C: Enterovirus. Pink line: TIANGEN, Purple line: Supplier A.
-
Compatible Instruments
● PRISM 7000/7700/7900HT, 7300/7500 Real-Time PCR System, 7500 Fast Real-Time PCR System, Viia 7 (Applied Biosystems)
● OPTICONTM/CFX96 (BIORAD)
● Light Cycler 480 (Roche)
● Smart Cycler® System (Cepheid)
● Mx3000P/Mx3005P (Stratagene)
● Other Real Time PCR thermal cycler
-
Sort by
-
Date
Date()
Date
Date()
Impact Factor
IF()
Impact Factor
IF()
