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What is Proteinase K? What Does Proteinase K Do in DNA Extraction?

Blog / 14/01/2024

Proteinase K is an important enzyme used in molecular biology labs for digesting and removing proteins from nucleic acid preparations. […]

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Strewbarries RNA Extraction to Identify Six Virus Using Multiplex PCR Assay Method

Blog / 23/01/2024

Background Strawberries are a type of perennial herbaceous plant in the Rose family (Rosaceae) and the Strawberry genus (Fragaria). They

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How To Improve the Efficiency of Crude DNA Extraction: Strategies and Optimization

Blog / 23/01/2024

Experiment design The experiment involves grinding and purifying plant materials to obtain a crude DNA extract. This extract is then

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What Does DNA Polymerase Do? Things you should know

Blog / 11/01/2024

DNA polymerase is an enzyme that plays an important role in DNA replication and is essential for life processes. Without

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What does RNase Do? Things you should know about

Blog / 08/03/2024

Ribonuclease, more commonly known as RNase, is an extremely important enzyme that plays a vital role in RNA metabolism within

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Conclusion Currently, gene expression studies mainly rely on qRT-PCR technology, commonly using reference genes to correct for errors introduced during sample detection. Earlier research generally assumed that housekeeping genes essential for maintaining cellular activities exhibit stable expression patterns under different conditions, being regarded as universal reference genes. However, with the advancement of high-throughput sequencing, more studies suggest that housekeeping genes are not always stable. Optimal reference genes often vary significantly among different species under different conditions. When studying the expression of target genes, using previously published reference genes without validation can lead to biased results. Hence, it's essential to reselect reference genes. In the case of the Cowpea weevil, it was found that the pairwise variation value Vn/n+1 in the GeNorm software's pairwise difference analysis was consistently less than the default value of 0.15 across different developmental stages and various tissues of adult weevils. Therefore, there is no need to include a third reference gene for data correction; the most suitable number of reference genes is two. Comprehensive assessment through △Ct, GeNorm, NormFinder, BestKeeper, and RefFinder online tool suggests using β-Act and β-Tub as internal reference genes for gene expression studies across different developmental stages of the Cowpea weevil. Similarly, for various tissues in adult weevils, β-Tub and α-Tub were recommended as internal reference genes. RPL40 was evaluated as relatively suitable for use as one of the internal reference genes in different tissues of adult Cowpea weevils. However, it appeared to be one of the least stable genes among different developmental stages of the weevils. On the other hand, GAPDH showed the least stability in expression among different tissues of adult Cowpea weevils and was also unsuitable as a reference gene for gene expression studies in different developmental stages of the weevils. Similar results were observed in other Coleopteran insects such as Colaphellus bowringi and Sympiezomias velatus (Tan et al., 2015; Li et al., 2018). This study provides accurate and reliable internal reference genes for accurately analyzing gene expression levels in different developmental stages and various tissues of the Cowpea weevil using qRT-PCR. Namun, whether these findings apply to other experimental conditions requires further investigation. The expression of internal reference genes in the Cowpea weevil varies significantly under different experimental conditions. This study utilized five methods to evaluate the expression stability of eight candidate internal reference genes in different developmental stages and various tissues of the Cowpea weevil. The results suggest that GeNorm software calculated V2/3 gene pairwise difference values under different conditions, all of which were below the threshold of 0.15. Karena itu, the optimal number of reference genes for studying relative gene expression is two. Specifically, for different developmental stages of the Cowpea weevil, it is recommended to use β-Act and β-Tub as internal reference genes. Meanwhile, for various tissues in adult weevils, β-Tub and α-Tub are recommended as internal reference genes.

Research on Selection of Reference Genes for Real-Time Fluorescent Quantitative PCR in Cowpea Weevils

Blog / 08/01/2024

About the research Currently used internal reference genes in qRT-PCR include various genes like ubiquitin-conjugating enzyme (UBC), ribosomal RNA (rRNA),

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The Nucleic Acid Testing Industry Expecting Growth is 6.8% in From 2023 ke 2033

Blog / 05/01/2024

The Nucleic Acid Testing Industry: Rapid Growth Driven by Technological Advances and Increasing Demand for Precision Diagnostics Nucleic acid testing

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How to conduct a DNA test to identify the male and female pigeons? Here is the most detailed process

Blog / 31/12/2023

Introduction Compared to conventional sex identification methods for birds, such as cloaca inspection, laparoscopy, steroid analysis, and karyotyping, PCR-based sex

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