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Analysis of var Gene Transcript Patterns by Quantitative Real-Time PCR

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Quantitative real-time PCR (qPCR) is a simple and sensitive method for determining the amount of a specific target DNA sequence present in a sample. Compared to RNA-seq, reverse transcription qPCR (RT-qPCR) is fast, requires only low input material and is easy to analyze. Therefore, qPCR is widely used to analyze gene expression in P. falciparum, including analyses of the multicopy gene families encoding variant surface antigens (VSAs), whose expression is clonally variant and prone to changes over time. In the recent years, several P. falciparum genomes of culture-adapted strains have been sequenced, providing the knowledge to design variable gene family-specific qPCR primers for each P. falciparum genetic background. Here, we describe the required materials, methods and key factors to perform RT-qPCR experiments to determine VSA transcript abundances in the P. falciparum clones 3D7/NF54, IT4, HB3, and 7G8.

OriginalsprogEngelsk
TidsskriftMethods in Molecular Biology
Vol/bind2470
Sider (fra-til)149-171
Antal sider23
ISSN1064-3745
DOI
StatusUdgivet - 2022

Bibliografisk note

© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

ID: 79741520