Product Information
GeneCopoeia’s ExoCt™ RT-qPCR System allows you to analyze gene expression by real-time qPCR directly from exosomes without any RNA purification. The ExoCt™ First-Strand cDNA Synthesis Kit contains ExoCt™ RT-for-All™ Buffer and ExoCt™ PAP/RTase Mix, which were specifically developed for robust first-strand cDNA synthesis covering a variety of RNA templates, including miRNA, mRNA, LncRNA, etc., from exosome lysates in a single reaction. The ExoCt™ SYBR® Green RT-qPCR Kit includes both RT and qPCR reagents, which combines PCR technology and SYBR® Green to make fast and accurate quantification of exosome RNAs. The system includes the ExoCt™ First-Strand cDNA Synthesis Kit and the ExoCt™ SYBR® Green RT-qPCR Kit.
Advantages
- Direct detection of gene expression from exosomes without any RNA purification
- Convenient reverse transcription of all exosomal miRNA, mRNA, and LncRNA in one reaction simultaneously
- Flexible analysis and quantitation of different kinds of RNA in downstream qPCR reactions
Simple Workflow
Isolated exosomes are lysed by adding ExoCt™ Lysis Buffer, then heating at 37°C for 10 min and 75°C for 10 min. Exosome lysates are used directly for ExoCt™ RT reactions, and the cDNA products can be used for qPCR to detect miRNA, mRNA, and LncRNA, etc.
Performance Data
Figure 1. Comparing RT-qPCR results from exosome lysates and exosome-extracted RNA. 3 x 107 cells were cultured in 175mm plate until reaching 70% to 90% confluency. 40ml cells culture media were harvested and exosomes were isolated using ExoSure™ Exosome Isolation Kit. Exosomes from HEK393(red), MSC(grey), H2228(yellow), H3122(blue) were either lysed with GeneCopoeia’s ExoCt™ Lysis Buffer, or the RNA was extracted from them. Exosome lysates, or extracted RNA were reverse-trascribed to cDNA using ExoCt™ RT regent. Then the cDNA was used for gene expression detection.
Figure 2. Reverse transcribing miRNA, mRNA, and LncRNA simutaneouly in one tube using RT-For-All™ RT regent. 3 x 107 cells were cultured in 175mm plate until reaching 70% to 90% confluency. 40ml cells culture media were harvested and exosomes were isolated using ExoSure™ Exosome Isolation Kit. Exosomes were lysed with GeneCopoeia’s ExoCt™ Lysis Buffer. Exosome lysates were reverse transcribed to cDNA with ExoCt™ RT-For-All™ regent. The cDNA was used for gene expression detection. A. Exosome lysates from HEK293 cells. B. Exosome lysates from H3122 cells.
Figure 3. RT-qPCR detection using exosome lysates. 3 x 107 cells were cultured in 175mm plate until reaching 70% to 90% confluency. 40ml cells culture media were harvested and exosomes were isolated using ExoSure™ Exosome Isolation Kit. Exosomes were lysed with GeneCopoeia’s ExoCt™ Lysis Buffer. Exosome lysates were either reverse transcribed into cDNA with ExoCt™ RT-For-All™ regent or Takara RT regent. The cDNA was then used for gene expression detection. A. Exosome lysates from MSC cells. B. Exosome lysates from H3122 cells.
