3rd July 2024

qScript Ultra Flex Kit. Faster, longer, tougher next gen cDNA synthesis with flexible priming

  • Enhanced Thermostability – Engineered for RT activity up to 65°C, overcoming challenging secondary structures
  • Superior Speed & Ease of Use – 10 minute reaction time with fewer pipetting steps
  • Maximum Yield & Sensitivity – Improved yields with limiting or compromised samples
  • Ultimate Inhibitor Resistance – Overcome a wide array of PCR inhibitors (salt, heparin, hematin, etc.)
  • Longer Transcripts – Ability to reverse transcribe single targets up to 20 kb

qScript Ultra Flex Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention or treatment of a disease.

Description
qScript Ultra Flex Kit is easy-to-use and highly efficient for the synthesis of first-strand cDNA with flexible priming methods including oligo-dT, random hexamer and gene specific priming to reverse transcribe RNA to cDNA.

A key component is a novel, state-of-the-art, RNase H deficient reverse transcriptase that was engineered for improved thermostability, velocity, processivity and resistance to many common reaction inhibitors. qScript Ultra Flex Kit contains all required components for first-strand cDNA synthesis except RNA template and gene specific primers. The cDNA product is directly compatible with downstream 2-step RT-qPCR or RT-PCR procedures.

Enhanced Thermostability & Superior Speed
qScript Ultra Flex Kit has an optimal reaction temperature of 55°C and a 10-min reaction time, significantly reducing time to result while working through challenging secondary RNA structures. The lower Cq values achieved for the engineered qScript Ultra reverse transcriptase indicate more efficient cDNA synthesis at higher temperatures, critical for progressing through high GC regions and regions of RNA secondary structure.

  •  Fastest total reaction time of 15 minutes

  • Optimal reaction temperature of 55°C, with high RT activity up to 65°C

High Yield & Sensitivity

qScript Ultra Flex Kit provides higher yield across various RNA input amounts. The sensitivity of this kit allows for low input/challenging sample quantities to be detected while maintaining linearity in cDNA synthesis.

  •  In randomer-primed first-strand cDNA synthesis reactions, the qScript Ultra Flex Kit shows improved cDNA conversion and allows for detection of sequence from highly degraded RNA isolated from formalin-fixed liver tissue.

  • Linearity is maintained in the presence of low and challenging input types
  • An RNA range of 100 pg – 10 ng was tested with a reproducible limit of detection (what is the limit of detection??)

  • Reproducible low limit of detection and broad and linear dynamic detection (8 orders of magnitude) down to 100 fg
  • qScript Ultra Flex Dynamic Range. In an oligo(dT)-primed first strand cDNA reaction series, the qScript Ultra Flex Kit shows a reproducible low limit of detection and a broad and linear dynamic detection range over an RNA input range greater than 8 orders of magnitude. 6 replicates of each input quantity were reacted with qScript Ultra Flex and then 1/10-volume of the firststrand product was analyzed for beta-actin sequence using PerfeCTa SYBR Green FastMix.

Inhibitor Resistance
qScript Ultra Flex Kit can readily reverse transcribe RNA while overcoming common inhibitors found in blood, skin, plants, feces and RNA extraction carryover chemicals. Relative to alternative products, qScript Ultra Flex Kit yields high levels of cDNA, even in the presence of a wide range of inhibitors, which may be carried over due to RNA isolation from difficult or challenging sample matrices.

  •  Inhibitor Resistance Across Wide Range of Sample Types. The robust, engineered qScript Ultra reverse transcriptase allows for tolerance to many common reaction inhibitors. In standard, randomer-primed first-stand cDNA synthesis reactions, the qScript Ultra Flex kit maintains high levels of cDNA products in the presence of several inhibitors as indicated by the lack of large qPCR Cq shifts in the treated samples. Kits from other vendors were reacted with and without inhibitors according to the manufacturer recommended protocols and often showed Cq shifts, indicating reduced cDNA yields.

Long Amplification

qScript Ultra Flex Kit has the ability to reverse transcribe long RNAs up to 20 kb, which enables full-length transcript analysis and facilitates splice variant characterization, or applications such as transcriptome and RNA virus sequencing. When paired with Quantabio repliQa HiFi ToughMix, long transcripts of 12.5 kb and 17.7 kb were able to be quickly reverse transcribed in 10 minutes followed by fast PCR amplification in 54 and 84 minutes, respectively.

cDNA synthesis reactions were carried out to 55°C for 10 minutes, and a portion of the products amplified using repliQa HiFi ToughMix, a high-fidelity endpoint PCR. A: Various quantities of rat brain total RNA were used for GSP-primed first-strand cDNA synthesis, then a 12.5 kb portion of the rat Dynein mRNA was amplified and analyzed. PCR amplification using the repliQa HiFi ToughMix took only 54 minutes. B: Various quantities of human total RNA were used for GSP-primed first-strand cDNA synthesis, then a 17.7 kb portion of the human SYNE1 mRNA was amplified and analyzed. PCR amplification using the repliQa HiFi ToughMix took only 1 hour and 24 minutes.

  • Ability to reverse transcribe single targets up to 20 kb
  • Ultra Fast amplification when paired with the Quantabio repliQa HiFi ToughMix

Components

Storage & Handling:

  • Store components in a constant temperature freezer at -25°C to -15°C.
  • After thawing, mix thoroughly before use.
  • For lot specific expiry date, refer to package label, Certificate of Analysis or Product Specification Form.

Additional Resources: