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prostar RT-PCR Systems for Robust High-Fidelity RNA Amplification

polymerase mixture, which has been specially formulated to achieve the highest replication accuracy possible for a Taq-based blend. TaqPlus Precision DNA polymerase exhibits a lower error rate than Taq and other commercially-available Taq-based DNA polymerase mixtures.10,11

prostar Ultra HF RT-PCR System

Stratagenes ProSTAR RT-PCR systems use Moloney murine leukemia virus RT (MMLV-RT) for cDNA synthesis of specific target RNAs. The ProSTAR Ultra HF RT-PCR system includes all reagents necessary to carry out cDNA synthesis [MMLV-RT, first-strand buffer, dNTPs, oligo(dT), random primers, and RNase-free water] using total RNA or poly(A)+ RNA. The kit also includes all reagents necessary for subsequent, ultra high-fidelity PCR amplification with PfuTurbo DNA polymerase (PfuTurbo, 10X Ultra-HF PCR buffer, dNTPs). In this system, the cDNA synthesis reagents and protocol provide optimal performance using reaction volumes ranging from 10 to 50 l. Because the kits protocol is versatile, investigators can conserve RNA and cDNA synthesis reagents when a limited number of PCRs are performed (e.g., 10-l volumes) or can scale up the reverse transcription reaction (e.g., 50-l volumes) when multiple targets are amplified from the same cDNA synthesis reaction.

Figure 1

The ProSTAR Ultra HF RT-PCR system has been used to accurately amplify a broad range of target sizes from a variety of RNA sources, including human, mouse, yeast, and bacteriophage total RNA or poly(A)+-selected mRNA. High product yields were achieved for targets ranging in length from 270 bp to 7.6 kb (Figure 1, Panels A and B). In addition to long target capability, the ProSTAR Ultra HF RT-PCR system exhibits high sensitivity. RT-PCR products can be amplified from as little as 1 fg (low comp
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