FastStart Universal Probe Master (Rox)

Description

• General description: 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 nonspecifically bind. FastStart™ Taq DNA Polymerase is activated by removing the blocking groups at a high temperature (i.e., a preincubation step at +95°C).
• Application: FastStart™ Universal Probe Master (Rox) has been used:in TaqMan® quantitative real-time polymerase chain reaction (qRT-PCR) reactions for the quantification of endogenous miRNAs, such as MIR376B, MIR376A and MIR181A[1][2][3]in reverse transcriptase (RT-PCR) to study tumor necrosis factor (TNF) expression in whole synovial tissue of undifferentiated peripheral inflammatory arthritis (UPIA) patients[4] for the amplification and detection of any DNA or cDNA target, including those that are GC- or AT-rich by quantitativePCR[5][6] Combine this master mix with Transcriptor First Strand cDNA Synthesis Kit (Roche) to achieve excellent results in two-step qRT-PCR.[7]
• Features and Benefits: Increase qPCR sensitivity and specificity.Produce lower cycle threshold (Ct) values.Use the master mix with any probe-based assay.Achieve sensitive, specific results in assays with the Universal ProbeLibrary Probes or any other hydrolysis probe.Amplify and detect a broad range of DNA or cDNA targets.Amplify fragments up to 500 bp long, including those that are GC- or AT-rich.Visualize amplification products on agarose gels.Use robotic pipetting stations to set up qPCR reactions. Use a master mix that is stable at room temperature during extended reaction setup times.Prevent false positives resulting from carryover contamination.Use this dUTP-containing mix with Uracil-DNA Glycosylase to eliminate contaminating DNA carried over from previous PCR reactions.
• Components: FastStart Universal Probe Master (Rox), 2x concentrated master mix that contains FastStart Taq DNA Polymerase, Reaction Buffer, Nucleotides (dATP, dCTP, dGTP, dUTP), and a reference dye.
• Quality: Function test: Each lot is tested for performance in qPCR using three templates: a GC–rich template, an AT-rich template, and a long template (approximately 440 bp).
• Other Notes: For life science research only. Not for use in diagnostic procedures
• Legal Information: FastStart is a trademark of Roche