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Experiment Tags: Knock-in Cell Lines, Knock-in
Experiment Summary: Because the success of knock-in cell line construction is influenced by various factors, including the length of the knock-in gene fragment, homologous recombination efficiency, and cell line type, which can impact editing efficiency, this also makes the knock-in technology challenging. This article introduces the principles of constructing KI cell lines and the details that need to be paid attention to.
Experiment Tags: Mitochondrial membrane potential, ΔΨm, JC-1 assay, apoptosis, mitochondrial dysfunction, flow cytometry, fluorescence microscopy, cell biology protocol
Experiment Summary: This protocol describes a JC-1–based method for assessing mitochondrial membrane potential (ΔΨm) in cultured cells. By comparing red and green fluorescence signals generated by JC-1 aggregates and monomers, respectively, changes in ΔΨm can be detected as an early indicator of mitochondrial dysfunction and apoptosis. The procedure applies to both flow cytometry and fluorescence microscopy and includes detailed guidance on cell preparation, staining, data acquisition, and analysis.
Experiment Tags: PCR, Standard PCR
Experiment Summary: Taq polymerase is frequently used in the polymerase chain reaction (PCR), for it is able to withstand repeated heating to 95 °C without significant loss of activity, an efficient method for greatly amplifying the quantity of DNA templates.
Experiment Tags: RT-PCR, PCR
Experiment Summary: RNA is first reverse transcribed into cDNA using a reverse transcriptase as described here, the resulting cDNA is used as templates for subsequent PCR amplification using primers specific for one or more genes.
Experiment Tags: PCR Amplification, Probe Hybridization, ELISA Assay
Experiment Summary: This experiment describes the procedure of PCR-ELISA experiment based on immunological method to detect PCR products.
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