Total RNA was isolated from hearts and cardiac endothelial cells, sorted with CD31 MicroBeads (Miltenyi Biotec) from the mouse hearts using the Trizol reagent (Invitrogen). The RNA pellet was dissolved in diethyl pyrocarbonate-treated H2O. First-strand cDNA synthesis was performed with 0.5 μg of total RNA using oligo(dT) primers and Superscript III reverse transcriptase (Invitrogen). One μL of the reaction product was taken for real-time RT-PCR amplification (ABI Prism 7000, Applied Biosystems) using a commercial SYBR® Green kit (Eurogentec, Angers, France). Primer sequences are available on request. Expression of each gene was normalized to the respective Gapdh, Actb, and Rplp0 expression.
Sybr green kit
Manufactured by Eurogentec
Sourced in France, Belgium
The SYBR® Green kit is a reagent used in real-time PCR (Polymerase Chain Reaction) applications. It contains SYBR® Green I dye, which binds to double-stranded DNA, allowing for the detection and quantification of DNA amplification during the PCR process.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using sybr green kit
1
Isolation and Analysis of Cardiac RNA
2
Isolation and Quantification of Cellular and EV-RNA
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