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- The technique of fluorescence in situ hybridization (FISH) is described. This is another method for examining sequence complexity within a genome. In this method, a DNA sequence, such as a particular gene sequence, can be detected within an intact chromosome by using a DNA probe that is complementary to the sequence.For example, let’s consider the β-globin gene, which isfound on human chromosome 11. A probe complementary to theβ-globin gene binds to that gene and shows up as a brightly colored spot on human chromosome 11. In this way, researchers can detectwhere the β-globin gene is located within a set of chromosomes. Becausethe β-globin gene is unique and because human cells are diploid(i.e., have two copies of each chromosome), a FISH experimentshows two bright spots per cell; the probe binds to each copy ofchromosome 11. What would you expect to see if you used thefollowing types of probes?A. A probe complementary to the Alu sequenceB. A probe complementary to a tandem array near…When performing QPCR to study differential gene expression, what is the role of SYBR green? What is the most correct answer? SYBRgreen intercalates proportionally and efficiently with double stranded DNA and can therefore be used to quantify it SYBRgreen intercalates proportionally and efficiently with nucleic acids and can therefore be used to quantify them. SYBRgreen intercalates more efficiently and proportionately with single stranded DNA and can therefore be used to quantify cDNA (a direct product of mRNA). SYBRgreen intercalates proportionally efficiently with messenger RNA and can therefore be used to quantify it.Northern blotting, RT-PCR, and microarrays can be used to analyze gene expression. A lab studies yeast cells, comparing their growth in two different sugars, glucose and galactose. One student is comparing expression of the gene HMG2 under these two conditions. Which technique(s) could he use and why? Another student wants to compare expression of all the genes on chromosome 4, of which there are approximately 800. What technique(s) could she use and why?
- Evidence [see P. G. Shiels, A. J. Kind, K. H. Campbell, et al. (1999),“Analysis of telomere lengths in cloned sheep,” Nature 399, 316–317] suggests that Dolly may have been genetically older than heractual age. As mammals age, the chromosomes in somatic cells tendto shorten from the telomeres. Therefore, older individuals haveshorter chromosomes in their somatic cells than do younger ones.When researchers analyzed the chromosomes in the somatic cells of Dolly when she was about 3 years old, the lengths of her chromosomeswere consistent with those of a sheep that was significantlyolder, say, 9–10 years old. (Note: As described in the chapter, thesheep that donated the somatic cell that produced Dolly was 6 yearsold, and her mammary cells had been grown in culture for severalcell doublings before one of the cells was fused with an oocyte.)A. Suggest an explanation why Dolly’s chromosomes seemedolder than they should have been.B. Let’s suppose that a female sheep (like Dolly), which…Should the exposome field focus exclusively on measuring chemicals derived from exogebeous sources or should the field also evaluate the chemicals generated and processed from endogeneous sources? If a study of the human genome gave rise to genomics, does the human exposome give rise to a new field of exposomics?Explain one experimental strategy for determining the functional role of the mouse HoxD-3 gene.
- Briefly describe the method of chromatin immunoprecipitation sequencing (ChIP-Seq). How is it used to determine nucleosome positions within a genome?How the amplification of other genomic methods such as optical mapping helps to overcome the shortfalls in sequencing data to achieve a chromosome-level genome assembly?You are using Drosophila as a model system to investigate eye development. You do positional cloning to isolate a mutant gene called eyeless (eyl), which causes flies to develop with reduced sized eyes when homozygous. Using several DNA markers from the candidate interval as probes for colony hybridization to a CDNA library, you obtain a 10 Kb cDNA insert cloned into the unique EcoRI site of a 3.5 Kb plasmid vector. a. Briefly describe how would you use the Drosophila eyl cDNA clone to identify a clone from a mouse genomic DNA library that is homologous to the Drosophila eyl gene? b. You isolate a clone of mouse genomic DNA that appears to be homologous to the Drosophila eyl gene. After DNA sequencing of a small piece of the mouse genomic clone, you use the DNA sequence to search the mouse genome database. You find that there is a SNP locus (n1234) that does NOT cause an RFLP in this genomic clone. You have identified several breeding lines of mice, each having been bred for several…
- How DNA recombination is used in unique variable region coding?In an in situ hybridization experiment, what is the relationship betweenthe base sequence of the probe DNA and the site on the chromosomal DNA where the probe binds?What are the three key processes that Pmar-1 activates in the micromeres? Explain the importance of timing in these events and in their relationship to each other.