FastStart SYBR Green Master

Uses

The FastStart^? SYBR^? Green Master has been used in qPCR and two-step qRT-PC in the SYBR^? Green I detection format. It is also used:

• in quantitative polymerase chain reaction (qPCR) for adeno-associated virus (AAV) titre quantification
• in qPCR to determine the expression level of different Col6a1-3 genes relative to glyceraldehyde 3-phosphate dehydrogenase (GAPDH)
• in quantitative real-time polymerase chain reaction (qRT-PCR) of G protein-coupled estrogen receptor-1 (GPER) mRNA
• in qRT-PCR for gene expression studies

General Description

FastStart SYBR Green Master; Instructions For Use

FastStart^? SYBR^? Green Master is a ready-to-use hot start reaction mix without ROX for quantitative polymerase chain reaction (qPCR) and reverse transcription (RT)-qPCR on real-time PCR systems other than the LightCycler^? instruments. This master mix simplifies the preparation of reactions for DNA detection and analysis. In combination with a real-time PCR instrument, suitable PCR primers, and a hydrolysis probe, FastStart^? TaqMan^? Probe Master allows very sensitive detection and quantification of defined DNA sequences.
SYBR^? Green I is a DNA double-strand-specific dye. During each phase of DNA synthesis, the SYBR^? Green I dye, included in the reaction mix, binds to the amplified PCR products. The amplicon can be detected by its fluorescence.
Hot start protocols have been shown to significantly improve the specificity, sensitivity, and yield of PCR. Heat-labile blocking groups on some of the amino acid residues of FastStart^? Taq DNA Polymerase make the modified enzyme inactive at room temperature. Therefore, there is no elongation during the period when primers can non-specifically bind. The FastStart^? Taq DNA Polymerase is activated by removing the blocking groups at a high temperature (i.e., a pre-incubation step at +95°C).
The FastStart^? SYBR^? Green Master can be used for the amplification and detection of any DNA or cDNA target, including those that are GC- or AT-rich. However, you would need to adapt your detection protocol to the reaction conditions of the particular real-time PCR instrument used and design specific PCR primers for each target.
Combine this master mix with our Transcriptor First Strand cDNA Synthesis Kit to achieve excellent results in two-step qRT-PCR.