RT-PCR & cDNA Synthesis Products for Gene Expression
RT-PCR and cDNA synthesis products provide the reverse transcriptase enzymes, buffers, primers, and master mixes needed to convert RNA templates into complementary DNA (cDNA) for downstream PCR or qPCR analysis, available in one-step and two-step kit formats. M-MLV and AMV-derived reverse transcriptases are available as stand-alone enzymes or complete kits with oligo(dT) and random primers.
Academic researchers running gene expression studies benefit from MBP's range, covering both routine and difficult RNA templates. Request a quote by contacting customerservice@mbpinc.net
RT-PCR & cDNA Synthesis Products
What Are RT-PCR & cDNA Synthesis Products?
RT-PCR (reverse transcription PCR) and cDNA synthesis products convert an RNA template into complementary DNA (cDNA) using a reverse transcriptase enzyme, providing the DNA template required for subsequent PCR or qPCR amplification of RNA targets. Products in this category include stand-alone reverse transcriptase enzymes (M-MLV, AMV-derived), complete one-step and two-step RT-PCR kits, cDNA synthesis master mixes, and supporting reagents such as oligo(dT) primers, random primers, and RNase inhibitors. Choose a two-step kit for flexibility across multiple downstream PCR targets from one RNA sample; choose a one-step kit for high-throughput simplicity when amplifying a single target.
What you will find:
- Reverse transcriptase enzymes for first-strand cDNA synthesis, including HighTherm™ Reverse Transcriptase (Azura) and OneScript Plus Reverse Transcriptase
- First-strand cDNA synthesis kits for RNA-to-cDNA conversion, including AzuraQuant™, AzuraFlex™, Accuris qMAX, OneScript Plus, and FS01 systems
- One-step RT-PCR kits combining reverse transcription and PCR amplification in a single workflow, including Azura 1-Step Ultra, Accuris One-Step, and MegaFi One-Step RT-PCR Kits
- RT-qPCR and multiplex-compatible systems, including Accuris qMAX Probe One-Step Multiplex RT-qPCR Kit (No ROX)
- miRNA-focused cDNA synthesis solutions, including miRNA All-In-One cDNA Synthesis Kit
- Supporting RT reagents and master mixes, including OneScript Hot RT enzymes and RT master mix systems with gDNA removal options
How to Choose RT-PCR & cDNA Synthesis Products
One-Step vs. Two-Step Format
One-step RT-PCR combines reverse transcription and PCR in a single reaction tube and buffer using gene-specific primers, simplifying workflow and reducing contamination risk, but limiting analysis to a small number of genes per RNA sample. Two-step RT-PCR performs reverse transcription separately, producing a cDNA pool that can be used across multiple subsequent PCR reactions targeting different genes.
Reverse Transcriptase Enzyme Choice
AMV reverse transcriptase is recommended for one-step and two-step RT-PCR, RT-qPCR, and reverse transcription of RNAs under 5 kb, particularly where the template RNA has strong secondary structure. M-MLV reverse transcriptase RNase H-minus point mutants are preferred for reverse transcribing long RNAs for cDNA library construction, cDNA probe generation, and primer extension.
Priming Strategy
Oligo(dT) primers target the poly-A tail of mRNA, enriching for mRNA-derived cDNA. Random primers (commonly hexamers) prime throughout the RNA template regardless of sequence, useful for templates lacking a poly-A tail or for generating full-length cDNA coverage, including 5' regions.
Throughput and Workflow Fit
One-step RT-PCR allows easy processing of large sample numbers and is amenable to high-throughput applications, though it may be less sensitive than two-step approaches in some scenarios since the reaction conditions are a compromise between RT and PCR optimums.
RNA Quality and Genomic DNA Elimination
Some cDNA synthesis kits include a genomic DNA elimination step before reverse transcription, important for accurate gene expression quantification, where genomic DNA contamination could produce a false signal in downstream qPCR.
Specifications Context
In a two-step process, the first step synthesises first-strand cDNA using a reverse transcriptase such as M-MLV-RT, and the resulting cDNA pool can then be used in multiple separate PCR reactions targeting different genes -- a key advantage when a single RNA sample needs to be analysed for several targets. AMV reverse transcriptase's tolerance of strong RNA secondary structure makes it a common choice for difficult templates, while M-MLV RNase H-minus variants are the standard for full-length cDNA library construction. cDNA synthesis reagents typically require −20 °C storage; RNase inhibitor components and RNase-free water are recommended throughout RNA-handling workflows to prevent template degradation.
Contact the expert team at MBP today and get optimized RT-PCR & cDNA Synthesis Products for your lab.
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