Cells were washed with PBS three times and then lysed in RNA lysis buffer (Peqlab). Frozen tissue samples were transferred to the lysis buffer and were disrupted by mechanical homogenization on ice. Matrigel drops containing organoids were scratched from the dish and centrifuged at 4°C, 800 g, for 3 min. Lysis buffer was added to the pellet containing the organoids, followed by gentle vortexing for 1 min. Then, total RNA was extracted using the peqGOLD Total RNA purification kit (Peqlab) according to the manufacturer's instructions.
Peqgold total rna purification kit
Manufactured by Avantor
The PeqGOLD Total RNA purification kit is a lab equipment product designed for the extraction and purification of total RNA from a variety of sample types. It utilizes a silica-based membrane technology to provide efficient and reliable RNA isolation.
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2 protocols using peqgold total rna purification kit
1
Extracting Total RNA from Organoids
2
Total RNA extraction and RT-qPCR analysis
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Total RNA was extracted from cells and tissues using the peqGOLD Total RNA purification kit (Peqlab) according to the manufacturer’s instructions. RNA was reverse transcribed into cDNA using the SensiFASTTM cDNA Synthesis Kit (Bioline, London, UK). Real time quantitative PCR (RT-qPCR) was performed according to Analytik Jena innuMIX qPCR Master mix SyGreen protocol (Jena, Germany). Primers used for RT-qPCR are shown in Table 1 . mRNA expression was normalized using suitable housekeeping genes as indicated in Table 1 and in the figure legends.
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