5. You are following a family that has a reciprocal translocation, where a portion of one chromosome is exchanged for another, creating hybrid chromosomes. In some cases of chronic myelogenous leukemia, patients will have a translocation between chromosome 9 and 22, such that portions of chromosomes 9 and 22 are fused together. You are choosing between performing FISH and G-banding, which technique is best used to find this translocation, and why did you choose this technique?
6. What type of nucleotide is necessary for DNA sequencing? How is it different structurally from a deoxynucleotide, and why is this difference necessary for sequencing? Below is a Sequencing gel. Please write out the resulting sequence of the DNA molecule. Blue = G, Red C, T=Green, A = Yellow (Please see below for the gel).

Answers

Answer 1

The best technique to detect the translocation in the family with reciprocal translocation would be Fluorescence In Situ Hybridization (FISH).

FISH is specifically designed to detect chromosomal abnormalities and rearrangements, such as translocations. It uses fluorescently labeled DNA probes that can bind to specific target sequences on the chromosomes. In the case of the translocation between chromosomes 9 and 22, FISH probes can be designed to specifically bind to the hybrid chromosomes formed by the fusion of these two chromosomes. By visualizing the fluorescent signals under a microscope, FISH allows for the direct detection and localization of the translocation event.

The nucleotide necessary for DNA sequencing is a deoxynucleotide triphosphate (dNTP). Structurally, a deoxynucleotide consists of a deoxyribose sugar, a phosphate group, and one of the four nitrogenous bases: adenine (A), cytosine (C), guanine (G), or thymine (T). The key difference between a deoxynucleotide and a nucleotide used in RNA (ribonucleotide) is the absence of an oxygen atom on the 2' carbon of the sugar in deoxynucleotides. This difference makes deoxynucleotides more stable and less susceptible to degradation.

During DNA sequencing, the incorporation of dNTPs is crucial. Each dNTP is complementary to the template DNA strand at a specific position. The DNA polymerase enzyme incorporates the appropriate dNTPs according to the template sequence, and the sequencing reaction proceeds by terminating the DNA synthesis at different points. By using dideoxynucleotides (ddNTPs) that lack the 3'-OH group necessary for further DNA elongation, the resulting DNA fragments can be separated by size using gel electrophoresis, as shown in the sequencing gel provided. The sequence of the DNA molecule can be determined based on the order of the colored bands, with blue representing G, red representing C, green representing T, and yellow representing A.

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