[Molecular techniques for the study of neoplasms] [Spanish]
The techniques used to study molecular tumors must be able to detect the mechanisms which are altered in neoplasia. According to the basic genetic mechanisms of neoplastic disease, the most frequently used techniques are:
1. Alterations in genetic dosage (gain / loss): Karyotypes, fluorescence in situ hybridization (FISH acronym in English), loss of heterozygosity (English LOH), comparative genomic hybridization (CGH).
2. Gene fusion / rearrangements: Southern / restriction fragment length polymorphism (RFLP in English), FISH, polymerase chain reaction after reverse transcription (RT-PCR Torit), real-time PCR.
3. Microsatellite alterations: Quantitative PCR amplification of polymorphic regions identifying alleles and allelic bands extra Southern blot / RFLP
4. Point mutations and isolated nucleotide polymorphisms (SNPs in English): Screening Test (single stranded conformational polymorphism (SSCP) or denaturing gradient gel electrophoresis (DGGE); Specific tests such as RFLP (Southern / PCR), allele-specific hybridization, PCR primers and sequencing allele-specific
5. Gene expression modifications: in situ hybridization (RNA), Northern blot, RT-PCR (quantitative), microarrays (cDNA, oligonucleotides, fabric)
6. Epigenetic modifications: methylation level by RFLP (Southern / PCR) or sequencing after chemical treatment
7. Mitochondrial genetic alterations: We studied with the same techniques used for elanalisis the nuclear genetic material.
8. Telomerase activity and telomere length analysis: amplification protocol telomeres, telomerase expression and telomerase-associated RNA (immunohistochemistry / in situ hybridization), telomere length (PCR / Southern).