Cycle 2

Denature and anneal primers o o

Fig. 6.3 Simplified PCR schema

Double-stranded DNA is denatured to allow binding of specific oligonucleotides on either side of the region of interest. Taq DNA polymerase extends the oligonucleotides before the double-stranded molecules are denatured and the process is repeated.

Repeat x 30

Oligonucleotide primer of different sample types, for example fresh, unfixed, cryopre-served and formalin-fixed paraffin-embedded tissue as well as hematoxylin and eosin-stained and formalin-fixed tissue.

PCR may be used to detect the presence of chromosomal translocations. The most commonly investigated rearrangements include the t(9;22) chromosomal translocation in chronic myeloid leukemia (CML), t(1;19), found in a subset of pre-B-cell ALL, the t(14;18) found in 85% of follicular and 15% of diffuse large cell lymphomas, and several others. Alternatively, in the lymphoid malignancies, if the tumor being investigated does not carry a translocation marker, PCR may be used to amplify rearranged antigen receptor [Ig or T-cell receptor (TCR)] genes.

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