Cells were grown in M199 media (Invitrogen, Grand Island, NY) supplemented with EGM-2 media (Invitrogen) at 5% CO2.
Extracted molecule
total RNA
Extraction protocol
Total RNA was extracted using Trizol Reagent Protocol.
Label
SYBER Green
Label protocol
PCR arrays were performed using the cell development & differentiation miRNA PCR array (SA Biosciences, Frederick, MD) following the Manufacturer's instructions. Reverse transcription was performed using 200ng of total RNA with the RT2 miRNA First Strand Kit (SA Biosciences). Quantitative real-time PCR were performed (IQ5, Bio-rad) with 40 cycles at 94oC for 15 seconds, 60oC for 60 seconds.
Hybridization protocol
n/a
Scan protocol
n/a
Description
Test
Data processing
Data normalization and analysis was performed according to the manufacturers instructions using their web-based software package: http://sabiosciences.com/pcrarraydataanalysis.php For normalization the average of 4 housekeeping genes (SNORD48, SNORD47, SNORD44, RNUG-2) was used. The sabiosciences web-based software performs all statistical analysis and deltadeltaCt based fold-change calculations. Fold change worksheet reports test/control (i.e. Cardiovascular progenitor cells/human embryonic stem cell) and test/test (i.e. adult cardiovascular progenitor cell clones/neonatal cardiovascular progenitor cell clones) ratios.
