FAQ’s

cDNA Synthesis

cDNA synthesis and RNA denaturation step
Contamination of RNA with RNase activity
Difference between qScript cDNA SuperMix and qScript cDNA Synthesis
Primers in qScript™ cDNA SuperMix
Priming strategies for first strand synthesis
Purpose of 25C incubation during cDNA synthesis
qScript™ Flex cDNA Kit vs qScript™ cDNA Synthesis Kit
RNA dynamic range with cDNA Supermix and cDNA synthesis Kit
RNA input for qScript cDNA Supermix
RNA input for qScript cDNA Synthesis Kit
Rnase Inhibitor and cDNA synthesis
scaling up a cDNA synthesis reaction
Stability of MMLV RT in qScript cDNA Supermix and qScript cDNA synthesis
Variability in the efficiency of first strand cDNA synthesis
What is the smallest quantity of RNA detectable by the qScript™ cDNA Synthesis Kit?

PCR

AccuFast and AccuStart Taq DNA Polymerase Activation time
AccuStart PCR SuperMix compatibility with T/A cloning methods
Effect of uracil residues on post PCR hybridization
Improving PCR specificity
Melting temperature (TM) of PCR product
Online registration when contacting technical support
PCR extension temperature
Product shelf life and storage
Reagents protocols, MSDS, PPS
RNA extraction methods
Volume of first strand reaction transferred to PCR

PCR Enzymes

AccuStart™ Taq DNA Polymerase proofreading activity
Activation of AccuStart™ Taq DNA Polymerase
Components of AccuStart™ Taq DNA Polymerase PCR buffer
Difference between Taq DNA Polymerase and AccuStart™ Taq DNA Polymerase
Function of the antibody in AccuStart™ Taq DNA Polymerase
Hot Start PCR
Inactivation of reverse transcriptase (RT)
Optimal Temperature for MMLV reverse transcriptase
Storage and stability of AccuStart™ Taq DNA Polymerase
Storage and stability of M-MLV reverse transcriptase (RT)
Unit definition for AccuStart™ Taq DNA Polymerase
Units of AccuStart™ Taq DNA Polymerase used in PCR
What is Uracil N Glycosylase (UNG)?

qPCR

Absolute versus relative quantification
Advantages of PerfeCTa™ qPCR FastMix™over PerfeCTa™ qPCR SuperMix
Applications for the qPCR FastMix
cDNA input in SYBR and Taqman qPCR
Compatibility of Quanta Biosciences reagents with Roche’s LightCycler 1.5/2.0
Difference between AccuFast™ Taq DNA polymerase and AccuStart™ Taq DNA Polymerase
Expiration date
Fast cycling reaction volume
Improving qPCR specificity
Information to provide when calling technical support
Instrument Compatibility- SYBR Green Detection
Instrument Compatibility-Taqman Probe Detection
Is the PerfeCTa™ qPCR FastMix™ offered with UNG?
Mixing PCR reactions
Multiplexing
One-step or Two-step reaction?
Performance of PerfeCTa™ qPCR FastMix compared with PerfeCTa™ qPCR Supermix
Primer quality/sequence effect on qPCR effeciency
Primer validation/quality
Replicates
Sensitivity of SYBR and Taqman reagents chemistries
Stability at room temperature
Using PerfeCTa™ qPCR SuperMix in Fast mode
What component in the qPCR FastMix allows rapid amplification results?

Probe Detection

Primer and probe concentration in a Taqman reaction

SYBR Detection

SYBR and ABI 7500 instrument

qRT-PCR

Dnase treatment of RNA
How to avoid Dnase treatment?
Inactivation of DNase during Dnase treatment compromises the quality of RNA
Is my RT-PCR product RNA specific?
qRT-PCR kits compatibility with Cell Lysis reagents
qRT-PCR Kits directly from cell lysates
Residual Dnase activity
RNA input in SYBR and Taqman qPCR
Setting up a no-RT (no-reverse transcription) control
Uracil N-glycosylase (UNG) and Reverse transcription