Assays to confirm viability: Immunohistochemistry with muscle specific antibodies
Growth protocol
Growth medium: HAM F/O Media supplements: 20% fetal calf serum, 4ng/ml bFGF Culture conditions: Collagen-coated plates
Extracted molecule
total RNA
Extraction protocol
RNA prep method: TriZol Cell purification method: Preplating method and serial passage Method for estimating purity: Desmin, Myf5, MyoD expressions
Label
Biotin
Description
This sample was analyzed as part of the Stem Cell Genomics Project (http://www.scgp.ca:8080/StemBase/). The biological material was submitted to the Ontario Genomics Innovation Centre (http://www.ottawagenomecenter.ca/) by Dr. Michael Rudnicki (mrudnicki@ohri.ca; 501 Smyth Road) for analysis. Stembase Experiment ID: E59 Stembase Experiment ID link: http://www.scgp.ca:8080/StemBase/?path=/browse/experiment&id=59 SCGP Sample ID: S78 SCGP Sample ID link: http://www.scgp.ca:8080/StemBase/?path=/browse/experiment&id=59#SAMPLE_59 Short description: Wild type myoblast A-C: Muscle satellite cells are a self-renewing pool of stem cells that give rise to daughter myogenic precursor cells in adult skeletal muscle. Published and preliminary data indicated that MyoD and p53 genes are involved in satellite cell differentiation. We would like to know what downstream genes of both transcription factors are affected in satellite cell-derived myoblasts (MyoD-/-, p53 -/-). Estimated purity: > 98% RNA concentration: 2.5 ug/ul Num cells for RNA prep: Growing myoblasts Sample volume: 10 ul
Data processing
Calculated using the MAS5 algorithm where sc=1500, tau=0.015, alpha1=0.04, and alpla2=0.06
