[NIPPON GENE] Real-time PCR Reagents
Real-time PCR is a method that uses the polymerase chain reaction (PCR) on a sample to determine the amount of DNA/RNA with a specific sequence. Use of this method allows determination of the expression level of the target gene or the content of viral genes in a sample. "qPCR" and "real-time PCR" are used as synonyms, because qPCR measurements are generally performed in real time. Two methods are often used: SYBR® Green and Fluorescent labeled probe.
* SYBR® is a registered trademark of Life Technologies
GeneAce qPCR Mix Series
▾GeneAce SYBR® qPCR Mix α
GeneAce SYBR® qPCR Mix α is a real-time PCR master mix (2 × concentration) for SYBR® Green detection systems compatible with real-time PCR thermal cyclers (plate type).
▾GeneAce Probe qPCR Mix II
GeneAce Probe qPCR Mix II is a real-time PCR master mix (2 × concentration) for fluorescent labeled probe detection systems compatible with real-time PCR thermal cyclers (plate type).
▾ Uracil-DNA Glycosylase (UNG)
UNG is available for the prevention of carry-over with PCR. UNG treatment before PCR can break down PCR products (including uracil residue) mixed in from previous reactions.
- High specificity and amplification efficiency
- Can be used for SNP genotyping experiments (probe detection systems)
- Can prevent carry-over with optional UNG addition (probe detection systems)
|Product Name||Volume||Available Instruments|
|SYBR®Green detection systems|
|GeneAce SYBR®qPCR Mix α||300 reactions※1||①|
|GeneAce SYBR®qPCR Mix α Low ROX||300 reactions※1||②|
|GeneAce SYBR®qPCR Mix α No ROX||300 reactions※1||③|
|Fluorescent labeled probe detection systems|
|GeneAce Probe qPCR Mix Ⅱ||200 reactions※2||①②③|
|Fluorescent labeled probe detection systems (carry-over prevention)|
|Uracil-DNA Glycosylase (UNG)||100 ng||-|
|GeneAce Probe qPCR Mix II with UNG||1 Set※3||①②③|
*1 300 reactions for 50 μL reaction systems, 750 reactions for 20 μL reaction systems
*2 200 reactions for 50 μL reaction systems, 500 reactions for 20 μL reaction systems
*3 A combination of GeneAce Probe qPCR Mix II (for 200 reactions) and UNG (100 ng)
① Devices that use high-concentration ROX for normalization (High ROX)
- ABI GeneAmp® 5700 ・ABI PrismTM 7000/7300/7700/7900HT ・MasterCycler® ep realplex
② Devices that use low-concentration ROX for normalization (Low ROX)
- ABI - Prism® 7500 ・ABI - QuantStudio® ・ABI - ViiATM 7 ・Stratagene/Agilebt - Mx3000P/3005P/4000
③ Devices that do not require ROX normalization (No ROX)
- BioRad - CFX384/CFX96 ・BioRad - DNA Engine Opticon® 1&2 ・BioRad - iCycler/iQ 5/My iQ/Chromo4
- Roche - LightCycler® 96/480/Nano ・Qiagen - Rotor-Gene® 2000/3000/6000/Q
- TaKaRa Bio - Thermal Cycler Dice®
*Please inquire about other real-time PCR devices.
DirectAce qPCR Mix plus ROX Tube
DirectAce qPCR Mix plus ROX Tube is a real-time PCR master mix (2 × concentration) for fluorescent labeled probe detection systems compatible with real-time PCR thermal cyclers (plate type), which can use DNA solutions crudely extracted from foods, etc. as template DNA solutions. The reagents are dispensed in small portions, facilitating ease of operation. 50 × ROX Passive Reference is attached to support devices that require ROX normalization.
This product is based on Ampdirect®(Shimadzu Corporation) and is optimized by combining it with a dedicated, hot-start PCR enzyme. It suppresses the effects of PCR inhibitors, allowing fluorescent labeled probe assays of various biological samples without DNA purification.
* Ampdirect® is a trademark of Shimadzu Corporation.
- Allows direct real-time PCR using crudely extracted DNA solutions as templates
- Can amplify the product even if sodium dodecyl sulfate (SDS), a PCR inhibitor, is present (0.1%) in the reaction mixture
- Separately attached ROX Passive Reference to support various devices
- Subdivided premix-type reagent for ease of operation
This product is not suitable for use with a TaqMan® MGB probe, which has a MGB (Minor Groove Binder) structure, as a fluorescent labeled probe.
This product does not support the standard protocol (95°C for 20 sec → [95°C for 3 sec → 60°C for 30 sec] × cycles) of the “Fast” run mode in real-time PCR thermal cyclers.
This product does not allow carryover treatment with UNG because it does not contain dUTP.
In Japan, DirectAce qPCR Mix plus ROX Tube is listed in the Grain Test and Grouping Test for Corn in the Consumer Affairs Agency "Annex: Examination Methods for Genetically Modified Foods after Safety Review." (as of June 2017)
GeneAce RL qPCR Mix with UNG
GeneAce RL qPCR Mix with UNG is a real-time PCR reagent for the intercalator method. If it is used with a real-time PCR instruments that requires ROX normalization, use of the separately available 50 × ROX Passive Reference is recommended.
- Highly sensitive detection of fluorescent signals
- Carryover treatment with UNG to prevent cross contamination
- Use of a hot start PCR enzyme to allow highly specific PCR
As it uses a hot-start PCR enzyme with chemical modification, enzymatic activity will not be sufficient without an enzymatic activation step (95°C, 10 min) and amplification efficiency may be reduced.
Other real-time PCR reagents
NIPPON GENE has a lineup of reagents for use in data correction or evaluation of real-time quantitative PCR.
|50× ROX Passive Reference||• Can be used with real-time PCR devices that require ROX correction|
|HEX Calibration Probe||• If HEX dye is not registered, calibration can be performed using this product|
|Internal Positive Control||• Can be used to evaluate the effect of false-negative results due to PCR inhibition
・Quality controlled product at approximately 20 copies/µL by digital PCR (measured values are within the range of 17 to 23 copies/µL)
・Can be used for genetic testing of various organisms, using artificial base sequences for detection
・Detection primers and probes are as follows:
IPC1-5’: 5’- CCG AGC TTA CAA GGC AGG TT-3’
IPC1-3’: 5’- TGG CTC GTA CAC CAG CAT ACT AG-3’
IPC1-Taq: 5’-(FAM) TAG CTT CAA GCA TCT GGC TGT CGG C (TAMRA)-3’
|GeneAce Reverse Trancriptase||• Modified (RNase H-) reverse transcriptase from M-MLV
• Improved synthetic efficiency for full-length cDNA
• Improved thermal stability
• High temperature reaction to avoid the effects of secondary structure of RNA
• Ideal for combination with GeneAce qPCR Mix α
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Real-time PCR Reagents -SYBR® Green Detection Systems-
Real-time PCR Reagents -Fluorescent labeled Probe Detection Systems-
Real-time PCR Reagents -Fluorescent labeled Probe Detection Systems- (carry-over prevention)
Real-time PCR Reagents -Fluorescent labeled Probe Detection Systems- (for Crude Samples)
Real-time PCR Reagents -Intercalator-
Other real-time PCR reagents
For research use or further manufacturing use only. Not for use in diagnostic procedures.
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