Analysis of telomeres and telomerase.
This unit describes techniques to analyze telomeric length and telomerase activity in human cells. Telomere length can be determined by a modification of Southern blotting in which the analysis of chromosome terminal restriction fragments (TRFs) provides the average lengths of all telomeres in a cell population. Telomerase activity can be measured in vitro by a sensitive and efficient polymerase chain reaction (PCR)-based detection method, also known as telomeric repeat amplification protocol (TRAP). These assays can be used to study the in vitro cellular effects of aging and cancer treatments on telomere biology and telomerase activity.
Herbert
Brittney-Shea
BS
The University of Texas Southwestern Medical Center, Dallas, Texas, USA.
Shay
Jerry W
JW
Wright
Woodring E
WE
D001483:Base Sequence; D015139:Blotting, Southern / Q000379:methods*; D045744:Cell Line, Tumor; D017931:DNA Primers / Q000378:metabolism; D004587:Electrophoresis, Agar Gel; D006801:Humans; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D011494:Protein Kinases / Q000378:metabolism; D019098:Telomerase / Q000378:metabolism*; D016615:Telomere / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism*
D017931:DNA Primers; D011494:Protein Kinases; D019098:Telomerase