dnasequencing.ppt

* Figure 8-35. In situ hybridization to locate specific genes on chromosomes. Here, six different DNA probes have been used to mark the location of their respective nucleotide sequences on human chromosome 5 at metaphase. The probes have been chemically labeled and detected with fluorescent antibodies. Both copies of chromosome 5 are shown, aligned side by side. Each probe produces two dots on each chromosome, since a metaphase chromosome has replicated its DNA and therefore contains two identical DNA helices. (Courtesy of David C. Ward.) * Figure 8-73. Using DNA microarrays to monitor the expression of thousands of genes simultaneously. To prepare the microarray, DNA fragments - each corresponding to a gene - are spotted onto a slide by a robot. Prepared arrays are also available commercially. In this example, mRNA is collected from two different cell samples for a direct comparison of their relative levels of gene expression. These samples are converted to cDNA and labeled, one with a red fluorochrome, the other with a green fluorochrome. The labeled samples are mixed and then allowed to hybridize to the microarray. After incubation, the array is washed and the fluorescence scanned. In the portion of a microarray shown, which represents the expression of 110 yeast genes, red spots indicate that the gene in sample 1 is expressed at a higher level than the corresponding gene in sample 2; green spots indicate that expression of the gene is higher in sample 2 than in sample 1. Yellow spots reveal genes that are expressed at equal levels in both cell samples. Dark spots indicate little or no expression in either sample of the gene whose fragment is located at that position in the array. (Microarray courtesy of J.L. DeRisi et al., Science 278:680686, 1997. With permission from AAAS.) * Figure 8-31 The DNA nucleotide sequences recognized by four widely used restriction nucleases. As in the examples shown, such sequences are often six base pairs long and palindromic (that