Home -> qPCR: SYBR® Green I Detection
SYBR® Green I Detection
Standard Cycling
PerfeCTa™ SYBR® Green SuperMixes are ready-to-use 2x reaction mixes that contain all components , except primers and template for real-time PCR using SYBR® Green. PerfeCTa™ SYBR® Green SuperMixes enable you to achieve exceptional reliability, sensitivity and specificity in qPCR. These SuperMixes provide sensitive detection of cDNA and genomic targets over a broad dynamic range with a strong fluorescent signal. PerfeCTa™ SYBR® Green SuperMixes are available and optimized for all real-time PCR instrument platforms including those requiring normalization with ROX reference dye. See our Selection Guide for SYBR Green I Detection or go directly to the corresponding qPCR product list.
- Reduced nonspecific amplification - AccuStart™ Taq DNA polymerase provides a superior, antibody-mediated, hot-start
- Minimal optimization - pre-mixed formats require only the addition of primers and template and are pre-optimized for most instruments
- Decontamination option - available with UNG (uracil-N-glycosylase) for elimination of previously-amplified fragments
FAST Cycling
PerfeCTa™ SYBR® Green FastMixes™ are ready-to-use 2x reaction mixes that contain all components, except primers and template for real-time PCR using SYBR® Green. PerfeCTa™ SYBR® Green FastMixes enable you to achieve exceptional reliability, sensitivity and specificity in qPCR. These SuperMixes provide sensitive detection of cDNA and genomic targets over a broad dynamic range with a strong fluorescent signal. PerfeCTa™ SYBR® Green FastMixes™ are available and optimized for all real-time PCR instrument platforms including those requiring normalization with ROX reference dye. See our Selection Guide for SYBR Green I Detection or go directly to the corresponding qPCR product list.
- AccuFast™ Taq DNA polymerase delivers maximum PCR efficiency, sensitivity, specificity and robust fluorescent signal using fast, or conventional, cycling protocols for SYBR® Green qPCR
- Instant activation - at 95º C AccuFast™ Taq DNA polymerase provides rapid recovery of fully active, unmodified Taq DNA polymerase
- Rapid polymerization kinetics - Replication of fragments up to 200 bp is complete in less than 20s at 60ºC
- Results in less than 1 Hour
Performance Data
PerfeCTa™ SYBR® Green SuperMix Comparison to Power SYBR® Green PCR Master Mix (Applied BioSystems®), PKCA
Log-fold serial dilutions of qScript™ cDNA from HeLa cell total RNA (100 ng to 1 pg) were analyzed by SYBR Green qPCR following each manufacturers protocol. Averaged plots of triplicate qPCR reactions are shown. No amplification was observed from 1pg input with Power SYBR Green PCR Master Mix.
More Performance Data
- PerfeCTa™ SYBR® Green SuperMix Comparison to SYBR® GreenER™ qPCR SuperMix (Invitrogen), PKCA
- PerfeCTa™ SYBR® Green SuperMix Comparison SYBR® GreenER™ qPCR SuperMix (Invitrogen), GAPD
- PerfeCTa™ SYBR® Green SuperMix Comparison to QuantiTect™ SYBR® Green PCR Kit (Qiagen), GAPD
- PerfeCTa™ SYBR® Green SuperMix Comparison to MESA GREEN qPCR MasterMix (ADAR)
Effect of dUTP Substitution on SYBR® Green qPCR (PKCA)
104-bp fragment of protein kinase C, alpha (PKCA) was amplified from log-fold serial dilutions of qScript™-synthesized cDNA from HeLa cell total RNA (100 ng to 10 pg) in 25-uL volumes with PerfeCTa™ SYBR Green SuperMix (dTTP-containing master mix) or PerfeCTa™ SYBR Green SuperMix, UNG (dUTP-containing master mix). Averaged plots of quadruplicate qRT-PCR reactions are shown. Amplification was not observed in "no template control" reactions (NTCs). PCR efficiency and sensitivity were comparable for the two amplification reagents. Inset: Dissociation curve for dT-containing (blue) and dU-containing (orange) PKCA-specific PCR product. Substitution of dUTP for dTTP lowered PCR product Tm by 1.3oC.
More Performance Data
Fast-cycle SYBR® Green qPCR with equal performance to conventional SYBR Green protocols
RNA-specific adenosine deaminase (ADAR) was amplified from log-fold dilutions of total HeLa cell cDNA (100 ng to 10 pg) using PerfeCTa™ SYBR Green FastMix™, with fast cycling protocol, or PerfeCTa™ SYBR Green SuperMix, with conventional cycling conditions.
More Performance Data
- PerfeCTa SYBR Green FastMix comparison to SYBR GreenER qPCR SuperMix.
- PerfeCTa™ SYBR® Green FastMix™ Comparison to iQ™ SYBR Green SuperMix
- PerfeCTa™ SYBR® Green FastMix™ Comparison to SYBR PreMix Ex Taq™
- Quanta Fast Cycling vs. Quanta Standard Cycling
- PerfeCTa™ SYBR® Green FastMix™ Comparison to DyNAmo™ Flash SYBR Green PCR Kit
- PerfeCTa™ SYBR® Green FastMix™ Comparison to QuantiFast™ SYBR Green PCR Kit
- Faster, More Sensitive SYBR Green qPCR
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