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RFLP Method - Restriction Fragment Length Polymorphism
作者:佚名 来源:davidson.edu 时间:2008-6-6

    Genetic Mapping

    To calculate the genetic distance between to loci, you need to be able to observe recombination. Traditionally, this was performed by observing phenotypes but with RFLP analysis, it is possible to measure the genetic distance between two RFLP loci whether they are a part of genes or not.

    Let's look at a simple example in fruit flies. Two RFLP loci with two RFLP bands possible at each locus:

    These loci are located on the same chromosome for the female (left) and the male (right). The upper locus can produce two different bands called 1 and 3. The lower locus can produce bands called 2 or 4. The male is homozygous for band 1 at the upper locus and 2 for the lower locus. The female is heterozygous at both loci. Thier RFLP banding patterns can be seen on the Southern blot below:

    The male can only produce one type of gamete (1 and 2) but the female can produce four different gametes. Two of the possible four are called parental because they carry both RFLP bands from the same chromosome; 1 and 2 from the left chromosome or 3 and 4 from the right chromosome. The other two chromosomes are recombinant because recombination has occurred between the two loci and thus the RFLP bands are mixed so that 1 is now linked to 4 and 3 is linked to 2.

    When these two flies mate, the frequency of the four possible progeny can be measured and from this information, the genetic distance between the two RFLP loci (upper and lower) can be determined.

    In this example, 70% of the progeny were produce from parental genotype eggs and 30% were produced by recombinant genotype eggs. Therefore, these two RFLP loci are 30 centiMorgans apart from each other.

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