Analysis of Mutant Trichoderma Afroharzianum MEA-12 by RT-qPCR (CAT#: STEM-MT-0067-LGZ)

Introduction

Cellulase enzymes play a key role in converting cellulosic biomass into fermentable sugars for the production of chemicals and fuels, which are often produced by filamentous fungi. However, most of the filamentous fungi obtained through natural breeding have low secretion ability when producing cellulase, which is far from meeting the requirements of industrial production. Combining random mutagenesis with adaptive laboratory evolution (ALE) strategies is an effective way to increase enzyme production in fungi. In addition, real-time reverse transcription quantitative PCR (RT-qPCR) can be used to analyze the transcript levels of major cellulase genes and transcription factor genes in isolated mutants.

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Principle Applications Procedure Materials Notes Related Products

Principle

Quantitative reverse transcription PCR (RT-qPCR) is an experimental method applied to PCR experiments using RNA as the starting material. In this method, total or messenger RNA (mRNA) is first transcribed into complementary DNA (cDNA) by reverse transcriptase. Subsequently, qPCR reaction was performed using cDNA as template.

Applications

Mutant Analysis

Procedure

1. Sample processing and preparation of PCR reaction system.
2. Add the amplification template, cover the PCR reaction cover, mix well, centrifuge at low speed instantaneously, and transfer to the PCR instrument.
3. Set the program for PCR amplification.
4. Data analysis.

Materials

Sample: depends on the customer's analysis requirements

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