SMARTSCRIBE REVERSE TRANSCRIPTASE KITS

Procurement Summary

SMARTScribe™ Reverse Transcriptase, 400 reactions/kit, manufactured by Takara Bio USA, Inc. NAICS CODE: 325414 - Biological Product (Except Diagnostic Manufacturing) Salient Characteristics- A modified Moloney Murine Leukemia Virus Reverse Transcriptase that synthesizes full-length cDNA from rare or long transcripts. SMARTScribe RT preparations are exceptionally pure, with all contaminating nucleases removed. These specific reverse transcriptase have the ability to synthesize a complementary DNA because the proprietary reagents used in these kits allow for three enzymatic activities: (1) RNA-dependent DNA polymerase, (2) RNase H, and (3) DNA-dependent DNA polymerase. First, RNA-dependent DNA polymerase synthesizes a DNA strand complementary to the RNA template. Then RNase H removes the RNA strand from the RNA–DNA hybrid double helix. The VRC Virology Laboratory comprises the Virology, Humoral Immunology, and Structural Biology Sections and related Cores. The goal of the Virology Laboratory is to conduct research to understand better the cellular and molecular regulation of viral gene expression, viral entry into the host cell, optimization of immune responses to gene-based vaccination and correlates of immune protection, assessment of active and passive approaches for HIV-1 vaccine development, and to apply structural biology techniques toward rational vaccine design for emerging and re-emerging infectious diseases. The Vaccine Research Center is developing vaccines for HIV-1 and other viruses. Currently, SBS/SBIS is working on developing vaccines for SARS-CoV-2. Production of wild-type and mutant viral proteins and anti-viral antibodies is critical in vaccine development. Making specific immunogens as probes is critical to isolate antibodies and develop therapeutic agents in the Covid19 treatment. The overarching goal of the project is to evaluate the immune response to HIV vaccines that employ novel germline-targeting immunogens for raising broadly neutralizing antibodies. This is achieved through single-cell next-generation sequencing of RNA isolated from probe-specific sorted B cells. Critical components of this workflow include reverse transcription of the isolated RNA and subsequent targeted amplification for downstream sequence analysis. Cutting-edge techniques and technologies are utilized in this project, including Illumina’s state-of-the-art NextSeq 1000 and NextSeq 2000 sequencing instruments. Only reagents and consumables specified for these platforms should be applied for optimal results, and Illumina’s sequencing protocols specify use of Takara Bio’s reagents. Takara Bio’s SMARTScribe™ Reverse Transcriptase kit contains a high-performance enzyme that performs unbiased cDNA synthesis, allowing for amplification and library construction from any RNA transcript. It generates long, full-length cDNA while preserving the relative transcript proportions of the original RNA sample, particularly crucial when transcribing RNA from a single cell. Takara Bio’s SeqAmp™ DNA Polymerase is a high-fidelity PCR enzyme that is specially formulated for next-Generation sequencing and transcriptome analysis. The specificity of this enzyme is especially critical for analysis of the highly variable sequence regions of B cells, which is what is being analyzed in this project. These reagents are formulated and optimized by Takara Bio specifically for use with Illumina’s sequencing protocols and are only available through Takara Bio.

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