10. TaqMan® Reverse Transcriptase-Polymerase Chain Reaction Coupled With Capillary Electrophoresis for Quantification and Identification
of bcr-abl Transcript Type
| Abstract |
|
|
Real-time TaqMan® polymerase chain reaction (PCR) assays allow quantification of the initial amount of target in a specimen,
specifically, and reproducibly. The major limitation of TaqMan PCR assays is that they do not detect the size of the amplified
target sequence. TaqMan PCR coupled with capillary electrophoresis is an alternative approach that can be used to circumvent
this limitation. In this chatper, the utility of this approach in the identification and quantification of bcr-abl fusion transcripts produced as a result of t(9;22)(q34;q11) in chronic myelogenous leukemia is described. In this assay,
abl primer labeled at its 5′-end with the fluorescent dye NED® (Applied Biosystems [ABI], Foster City, CA) is incorporated into
the bcr-abl fusion product during the real-time PCR. The incorporated NED fluorescent dye is then used subsequently to identify the specific
fusion transcript present in a given specimen by high-resolution capillary electrophoresis and GeneScan® (ABI) analysis. Knowledge
of the type of fusion transcript present in a specimen is useful to rule out false-positive results and to compare clones
before and after therapy.
Series: Methods in Molecular Biology | Volume: 335 | Pub. Date: Apr-01-2006 | Page Range: 135-145 | DOI: 10.1385/1-59745-069-3:135
Subject: Biochemistry
Key Words: Real-time TaqMan PCR, chronic myelogenous leukemia -
bcr-abl fusion transcripts - abl - GeneScan - capillary electrophoresis
Comments (Loading...) |
||
Loading... |





















