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Codex® HiTemp Reverse Transcriptase
A reverse transcriptase designed for challenges

Codex® HiTemp Reverse Transcriptase is a highly sensitive and robust enzyme specifically engineered for thermostability and performance in challenging conditions. This RNase H minus enzyme is automation-friendly with extended stability at ambient temperatures. An expedited protocol saves considerable time to results while retaining key performance metrics such as sensitivity, making it the ideal choice for custom one-step RT-qPCR assays. Codex® HiTemp Reverse Transcriptase is available for bulk purchases as an OEM product. Customization options are available.

Improve detection of difficult targets
Thermostability up to 70°C

Codex® HiTemp Reverse Transcriptase is engineered for enhanced thermostability, performing reliably at temperatures as high as 70°C when tested across a panel of 30 genes. This allows for maximum success of first strand synthesis from a broad range of target RNA species with varied levels of secondary structures.

Automate one-step RT-qPCR assays
Stability at ambient temperature for at least 5 hours

When formulated with wild-type Taq polymerase and reaction buffer, Codex® HiTemp Reverse Transcriptase is stable for at least five hours at room temperature, making it amenable for automation applications.

Codex® HiTemp Reverse Transcriptase can perform at temperatures as high as 70°C across multiple gene targets

RT Fig 1 Temp

Figure 1. Codex® HiTemp Reverse Transcriptase can perform at temperatures as high as 70°C across multiple gene targets. A one-step RT-qPCR mix containing Codex® HiTemp Reverse Transcriptase and wild-type Taq polymerase was challenged by performing reverse transcription at 50°C, 60°C, and 70°C, for a panel of 30 unique genes. Codex® HiTemp Reverse Transcriptase was able to reliably detect all targets at each temperature tested.

Codex® HiTemp Reverse Transcriptase demonstrates high sensitivity and exceptional linearity in a 5-minute RT reaction

RT Fig 2 High Sensitivity

Figure 2. Codex® HiTemp Reverse Transcriptase demonstrates high sensitivity in a 5-minute RT reaction. Sensitivity and efficiency of a one-step RT-qPCR mix containing Codex® HiTemp Reverse Transcriptase and wild-type Taq polymerase was compared between a standard and expedited protocol across RNA inputs ranging from 100 ng to 0.1 pg. Reverse transcription was carried out in either 15 or 5 minutes, while the annealing-extension step during amplification was either 30 or 15 seconds per cycle respectively. Cq values were similar for both for each input amount. The expedited protocol retained sensitivity down to 0.1 pg input RNA.

Codex® HiTemp Reverse Transcriptase maintains performance with degraded RNA

RT Fig 3 Degraded RNA Full

Figure 3. Codex® HiTemp Reverse Transcriptase maintains performance with degraded RNA. The performance of a one-step RT-qPCR mix containing Codex® HiTemp Reverse Transcriptase and wild-type Taq polymerase was benchmarked against a commercially available one-step RT-qPCR mix (Supplier 1) using RNA with varying degrees of degradation as input. Both one-step RT-qPCR mixes performed similarly for intact and minimally degraded RNA samples (RQS values of 9.1 and 7.2, respectively). However, the Codex® HiTemp Reverse Transcriptase containing mix demonstrated improved performance over Supplier 1 when RNA samples with higher degrees of degradation were used. Here, the ΔCq values between intact and highly degraded RNA were as low as 0.54 cycles for the one-step RT-qPCR mix containing Codex® qPCR Reverse Transcriptase, compared to 0.90 cycles for Supplier 1.

Obtain reliable performance in challenging assay conditions:
Degraded RNA, wide range of pH, and common inhibitors

Codex® HiTemp Reverse Transcriptase performs reliably with degraded RNA, across a pH range of 8.0 to 8.8, and can retain activity in the presence of common inhibitors for unhindered performance.

Detect low levels of target RNA
Sensitivity down to 0.1 pg

Codex® HiTemp Reverse Transcriptase can detect target RNA from as little as 0.1 picograms total RNA. This optimum sensitivity minimizes failures of assays with low amounts of starting material.

Expedite RT-qPCR assay time
RT reaction in 5 min

Codex® HiTemp Reverse Transcriptase is capable of first strand synthesis in as little as five minutes while retaining sensitivity. In combination with a commercially available wild-type Taq polymerase, the subsequent PCR amplification has been demonstrated in a rapid protocol using an annealing-extension time of just 15 seconds instead of the standard 30 seconds per cycle, further reducing assay time significantly.

Multiplex with ease
Up to 4-plex demonstrated

Codex® HiTemp Reverse Transcriptase performs seamlessly in multiplex assays with up to four targets.

OEM enzyme supply

Codex® HiTemp Reverse Transcriptase is available for OEM supply. Purchase quantities and concentrations can be customized for seamless integration into one-step RT-qPCR assays.

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