Analysis of Transcriptional Level of the Pmrk And Pmrc Genes Encoding for Lipopolysaccharide Target Modifying Enzymes by RT-qPCR (CAT#: STEM-MT-0054-LGZ)

Introduction

Colistin is one of the last-line therapeutic agents against multidrug-resistant Gram-negative bacteria (GNB), including Klebsiella pneumoniae. To assess the relationship between upregulation of the pmrHFIJKLM and pmrCAB operons and colistin resistance, RT-qPCR analysis was used to quantify the transcript levels of the pmrK and pmrC genes encoding LPS target modifying enzymes.

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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

Gene Transcription Levels

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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