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TGGETemperature Gradient Gel Electrophoresis (nucleic acid analysis)
TGGEThermal Gradient Gel Electrophoresis
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Diagnosis (N = 118) SB TGGE T-cell neoplasms (n = 58) 58 54 CTCL (n = 11) PTCL (n = 40) LGL (n = 3) PLL (n = 2) PTLD (n = 2) B-cell neoplasms (n = 38) 0 1 CLL/SLL (n = 11) FL (n = 7) MCL (n = 2) MALToma (n = 4) LCL (n = 14) Reactive (n = 22) 0 0
SB column indicates TCR[Beta] gene rearrangement positivity as determined by Southern blotting using TCR[Beta] probe; TGGE, clonality as determined by polymerase chain reaction amplification of TCR[Gamma], region; CTCL, cutaneous T-cell lymphoma, including mycosis fungoides; PTCL, peripheral T-cell lymphoma; LGL, large granular cell leukemia; PLL, prolymphocytic leukemia; PTLD, posttransplant lymphoproliferative disorder; CLL/SLL, chronic lymphocytic leukemia/small lymphocytic lymphoma; FL, follicular lymphoma; MCL, mantle cell lymphoma; MALToma, mucosa-associated lymphoid tissue lymphoma; and LCL, large cell lymphoma.
Fifty-four (93%) of 58 cases of lymphomas with evidence of TCR[Beta] gene rearrangement based on Southern blot analysis showed rearrangement pattern by TGGE analysis of the TCR[Gamma]-chain gene (Figure 1).
We established a TGGE method to screen for sequence variations within the region of the apo B gene that included the putative receptor-binding domain of the molecule.
We chose TGGE because it affords a high sensitivity (>95%) for point mutations if experimental conditions are optimized.
We therefore strongly recommend the use of screening strategies, such as TGGE, capable of detecting all known functionally relevant sequence variations of the apo B-100 receptor-binding domain.
Comparative studies prove that TGGE [9] or DGGE [54] detects a higher proportion of point mutations than SSCP.
1) Nonstandard abbreviations: DGGE, TGGE, denaturing, temperature gradient gel electrophoresis; ds, double stranded; ss, single stranded; SSCP, single-strand conformation polymorphism; HET, heteroduplex analysis; CCM, chemical cleavage method; EMC, enzyme mismatch cleavage; CFLP, cleavage fragment length polymorphism; PTT, protein truncation test; SDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis; RT, reverse transcription; ASO, allele-specific oligonucleotide; RFLP, restriction fragment length polymorphism; PODGE, profiling of oligonucleotide dissociation gel electrophoresis; ASA, allele-specific amplification; ARMS, amplification refractory mutation system; ASPCR, allele-specific PCR; and OLA, oligonucleotide ligation assay.