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highQu® HighScriber™ Reverse Transcriptase Mix

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Product Description

The HighScriber™ Reverse Transcriptase Mix is a premium tool for the high efficiency reverse transcription of up to 12-15 kb long cDNA. Mix includes HighScriber™ Reverse Transcriptase and Ribonuclease Inhibitor for save, robust cDNA synthesis and ease of use. HighScriber™ Reverse Transcriptase allows for high detection sensitivity from 1 pg of total RNA. The wide reaction temperature range (38°C - 55°C) ensures efficient cDNA synthesis from complex or GC rich templates.

The enzyme uses ssRNA or ssDNA as a template, possesses no detectable Ribonuclease H activity specific to RNA in RNA-DNA hybrids. A highly reduced Ribonuclease H activity allows for transcription of full lengths long transcripts. HighScriber™ Reverse Transcriptase can be used for RACE as it has terminal transferase activity - adds cytosines to 3’ ends of cDNA.
The Ribonuclease inhibitor premixed with the RT ensures RNA protection from ribonuclease degradation.
Supplied 5X ALLin™ HighScriber Buffer includes everything you need for the cDNA synthesis reaction. To minimize pipetting steps it contains MgCl2, dNTPs, enhancers, stabilizers. The only things to add is the template RNA and primer.

Choosing the right reverse transcriptase

Different reverse transcriptase used for cDNA synthesis have different properties and therefore not all of them are suitable for the same applications. Here are some important factors to consider when choosing the right reverse transcriptase:

•          Some reverse transcriptases work at lower temperatures, such as 42°C (MMuLV RT), while thermostable reverse transcriptases work at higher temperatures, such as 50°C or 55°C (highQu HighScriber™ RT). The more complex templates are transcribed, the higher temperature is recommended to disrupt secondary structures of the RNA.

•          Reverse transcriptases might have template preferences and are more efficient at synthesizing cDNA from certain RNA templates, such as mRNA, while others can synthesize cDNA from a wider range of RNA templates.

•          The sensitivity of a reverse transcriptase becomes especially important if the RNA sample is of low copy-number concentration. Not all enzymes can synthesize cDNA from low amounts of RNA efficiently.

•          The speed of reverse transcriptases also differs. Some RT enzymes work faster than others and ensure high cDNA synthesis yield in less time.

Good enzymes are typically supplied with optimized reverse transcription reagents to ensure high reverse transcription yields, enable high temperature protocols, provide high specificity and unbiased cDNA synthesis.

Applications

  • cDNA synthesis of up to 15 kb long transcripts.

  • Template generation for RT-PCR & RT-qPCR.

  • cDNA synthesis from complex templates.

Benefits

  • Thermostable Reverse Transcriptase blended with Ribonuclease Inhibitor for efficient cDNA synthesis.

  • High yields of full lengths transcripts up to 12-15 kb.

  • cDNA synthesis from complex templates at up to 55°C.

  • High sensitivity detection from 1 pg of total RNA template.

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