An inexpensive gel electrophoresis-based polymerase chain reaction method for quantifying mRNA levels.
In order to engage their students in a core methodology of the new genomics era, an ever-increasing number of faculty at primarily undergraduate institutions are gaining access to microarray technology. Their students are conducting successful microarray experiments designed to address a variety of interesting questions. A next step in these teaching and research laboratory projects is often validation of the microarray data for individual selected genes. In the research community, this usually involves the use of real-time polymerase chain reaction (PCR), a technology that requires instrumentation and reagents that are prohibitively expensive for most undergraduate institutions. The results of a survey of faculty teaching undergraduates in classroom and research settings indicate a clear need for an alternative approach. We sought to develop an inexpensive and student-friendly gel electrophoresis-based PCR method for quantifying messenger RNA (mRNA) levels using undergraduate researchers as models for students in teaching and research laboratories. We compared the results for three selected genes measured by microarray analysis, real-time PCR, and the gel electrophoresis-based method. The data support the use of the gel electrophoresis-based method as an inexpensive, convenient, yet reliable alternative for quantifying mRNA levels in undergraduate laboratories.
Bradford
William D
WD
Missouri Western State College, Saint Joseph, MO 64507, USA.
Cahoon
Laty
L
Freel
Sara R
SR
Hoopes
Laura L Mays
LL
Eckdahl
Todd T
TT
D001695:Biology / Q000379:methods; D019411:Clinical Laboratory Techniques; D004504:Education, Medical, Undergraduate / Q000379:methods; D018961:Educational Technology; D004587:Electrophoresis, Agar Gel / Q000191:economics* / Q000379:methods*; D005821:Genetic Techniques; D016681:Genome, Fungal; D023281:Genomics / Q000379:methods; D020411:Oligonucleotide Array Sequence Analysis; D018794:Problem-Based Learning; D012333:RNA, Messenger / Q000032:analysis* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012441:Saccharomyces cerevisiae / Q000235:genetics; D013663:Teaching / Q000379:methods
D012333:RNA, Messenger