Quantitative real-time PCR expression analysis of peripheral blood mononuclear cells in pancreatic cancer patients.
The ability of peripheral blood mononuclear cells (PBMCs) to act as a surrogate window into the presence and physiologic effects of pancreatic cancer is becoming increasingly apparent. In this chapter, we describe the techniques for isolation, lysis, RNA extraction, cDNA synthesis, and Q-RT PCR analysis of PBMCs as well as reasonable alternatives and the advantages and disadvantages of each. We further discuss the noteworthy considerations necessary for successful isolation and conversion of the high-quality PBMC RNA required to acquire interpretable and reproducible results for PBMC genetic expression analysis.
Department of Biochemistry and Molecular Biology, Eppley Institute for Research in Cancer and Allied Diseases, Omaha, NE, USA.
Michael J
MJ
Baine
Kavita
K
Mallya
Surinder K
SK
Batra
D002469:Cell Separation / Q000379:methods; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000378:metabolism*; D010190:Pancreatic Neoplasms / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D013048:Specimen Handling