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A 13-nucleotide section of an autoradiogram from a Sanger sequencing experiment is depicted in the image below. Based on the band pattern observed here, write out the sequence of both the complementary strand generated during the experiment, and the template strand that is being analyzed. Be sure to clearly indicate the 5' and 3' ends of each. ddATP ddGTP ddCTP 1 ddTTP 3' 5' Tyne a short answer in the space provided below

A 13-nucleotide section of an autoradiogram from a Sanger sequencing experiment is depicted in the image below. Based on the band pattern observed here, write out the sequence of both the complementary strand generated during the experiment, and the template strand that is being analyzed. Be sure to clearly indicate the 5' and 3' ends of each. ddATP ddGTP ddCTP 1 ddTTP 3' 5' Tyne a short answer in the space provided below

Human Anatomy & Physiology (11th Edition)
Human Anatomy & Physiology (11th Edition)
11th Edition
ISBN:9780134580999
Author:Elaine N. Marieb, Katja N. Hoehn
Publisher:Elaine N. Marieb, Katja N. Hoehn
Chapter1: The Human Body: An Orientation
Section: Chapter Questions
Problem 1RQ: The correct sequence of levels forming the structural hierarchy is A. (a) organ, organ system,...
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**Sanger Sequencing Autoradiogram Analysis** In this exercise, a 13-nucleotide section of an autoradiogram from a Sanger sequencing experiment is provided. The goal is to interpret the band pattern and determine the sequence of both the complementary strand generated during the experiment and the original template strand being analyzed. **Band Patterns and ddNTP Incorporation:** The image displays a sequencing gel with four lanes, each labeled at the top with a specific dideoxynucleotide triphosphate (ddNTP): ddATP, ddGTP, ddCTP, and ddTTP. Each lane corresponds to a specific nucleotide terminator used in the sequencing reaction. **Description of the Gel Diagram:** - **Lane ddATP:** Contains bands at various positions. - **Lane ddGTP:** Contains bands at various positions. - **Lane ddCTP:** Contains bands at various positions. - **Lane ddTTP:** Contains bands at various positions. The bands in each lane represent DNA fragments that terminated at that specific ddNTP. Reading from the bottom (5' end) to the top (3' end) of the gel provides the sequence of the complementary strand in the 5’ to 3’ direction. **Interpreting the Sequences:** 1. **Complementary Strand Sequence (from Gel):** - Read the sequence in the 5’ to 3’ direction from the bottom to the top of the gel by locating the position of each band and noting the corresponding ddNTP. 2. **Template Strand Sequence:** - Once the complementary strand sequence is determined, calculate the template strand by converting each base to its complement (A↔T, C↔G). Be sure when completing the exercise to clearly indicate the 5’ and 3’ ends of both strands. This understanding is crucial for determining the correct DNA sequence of the original sample.
Transcribed Image Text:**Sanger Sequencing Autoradiogram Analysis** In this exercise, a 13-nucleotide section of an autoradiogram from a Sanger sequencing experiment is provided. The goal is to interpret the band pattern and determine the sequence of both the complementary strand generated during the experiment and the original template strand being analyzed. **Band Patterns and ddNTP Incorporation:** The image displays a sequencing gel with four lanes, each labeled at the top with a specific dideoxynucleotide triphosphate (ddNTP): ddATP, ddGTP, ddCTP, and ddTTP. Each lane corresponds to a specific nucleotide terminator used in the sequencing reaction. **Description of the Gel Diagram:** - **Lane ddATP:** Contains bands at various positions. - **Lane ddGTP:** Contains bands at various positions. - **Lane ddCTP:** Contains bands at various positions. - **Lane ddTTP:** Contains bands at various positions. The bands in each lane represent DNA fragments that terminated at that specific ddNTP. Reading from the bottom (5' end) to the top (3' end) of the gel provides the sequence of the complementary strand in the 5’ to 3’ direction. **Interpreting the Sequences:** 1. **Complementary Strand Sequence (from Gel):** - Read the sequence in the 5’ to 3’ direction from the bottom to the top of the gel by locating the position of each band and noting the corresponding ddNTP. 2. **Template Strand Sequence:** - Once the complementary strand sequence is determined, calculate the template strand by converting each base to its complement (A↔T, C↔G). Be sure when completing the exercise to clearly indicate the 5’ and 3’ ends of both strands. This understanding is crucial for determining the correct DNA sequence of the original sample.
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