出生前基因型.pptVIP

Prenatal genotyping using RFLP The differences in RFLP patterns are used to diagnose genetic disorders prenatally and can also determine the genotype of parents and other family members who may be heterozygous carriers. For prenatal diagnosis fetal cells are obtained by amniocentesis The tools needed to perform these DNA diagnoses are first we need a restriction enzyme that will cut DNA in a manner that is informative for the disease in question, and second we need a nucleic acid probe that will detect that region 1kb 4kb sample A: probe xxxxxxxxxx Sample B : xxxxxxxxxxx The two DNA samples are cut with the appropriate enzyme and are hybridized with the correct probe. The probe hybridized to the 1 and 4 kb fragments in sample A and the 5 kb fragment in sample B. This is an example where a restriction site has been gained in sample A (or lost in sample B) by mutation. Sample A Sample B -------------------------- 5.0 kb ___ 4.0 kb ___ 1.0 kb ___ --------------------------- Genotype Determination using RFLPs and a Gene Probe A DNA probe that hybridizes to the 5 end of the human beta globin gene (shown in blue on the diagram below) was used to identify RFLP pieces from members of a family in which sickle-cell hemoglobin (HbS) was segregating. The normal HB allele (HbA) is cut at three places by a particular restriction enzyme (positions shown with red arrows). The HbS mutation destroys the internal restriction site so the HbS gene is cut in only two places. Thus, the probe hybridizes to a 1.15 kb DNA fragment from HbA DNA and hybridizes to a 1.35 kb fragment from HbS DNA. Linked DNA markers All of the DNA (RFLP or PCR) markers that we have discussed so far have been targeted to a specific gene. For many important traits, the actual gene of interest is not known. Therefore, probing for th