You want to study a biomolecule in the laboratory. You have ordered the synthetic gene from a company and want to check that it has been introduced into your model organism. What is the proper order of experiments below for doing this? I. Sanger sequencing II. Transformation III. Restriction digest IV. DNA purification from a single colony I, II, III, IV I, IV, II, III III, II, IV, I II, III, IV, I IV, II, III, I

Biochemistry
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ISBN:9781319114671
Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Chapter1: Biochemistry: An Evolving Science
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**Title: Proper Order of Experiments to Verify Gene Introduction in a Model Organism**

**Description:**

When studying a biomolecule in the laboratory and ordering a synthetic gene, it's crucial to verify its introduction into your model organism. The experiments should be conducted in the following order to ensure accuracy:

1. **Restriction Digest (III)**
   - Begin by using restriction enzymes to verify the presence of the gene by cutting the DNA at specific sites, allowing confirmation of correct gene insertion.

2. **Transformation (II)**
   - Introduce the synthetic gene into the host organism's cells, typically using methods such as heat shock or electroporation for bacterial cells.

3. **DNA Purification from a Single Colony (IV)**
   - Isolate and purify DNA from a single transformed colony to prepare it for sequencing analysis.

4. **Sanger Sequencing (I)**
   - Perform Sanger sequencing to determine the exact nucleotide sequence of the inserted gene, confirming its correct integration and sequence accuracy.

**Highlighted Correct Order:**
- The order highlighted in green indicates the proper sequence: **III, II, IV, I**

This sequence ensures that the gene is properly introduced and accurately verified within the model organism.
Transcribed Image Text:**Title: Proper Order of Experiments to Verify Gene Introduction in a Model Organism** **Description:** When studying a biomolecule in the laboratory and ordering a synthetic gene, it's crucial to verify its introduction into your model organism. The experiments should be conducted in the following order to ensure accuracy: 1. **Restriction Digest (III)** - Begin by using restriction enzymes to verify the presence of the gene by cutting the DNA at specific sites, allowing confirmation of correct gene insertion. 2. **Transformation (II)** - Introduce the synthetic gene into the host organism's cells, typically using methods such as heat shock or electroporation for bacterial cells. 3. **DNA Purification from a Single Colony (IV)** - Isolate and purify DNA from a single transformed colony to prepare it for sequencing analysis. 4. **Sanger Sequencing (I)** - Perform Sanger sequencing to determine the exact nucleotide sequence of the inserted gene, confirming its correct integration and sequence accuracy. **Highlighted Correct Order:** - The order highlighted in green indicates the proper sequence: **III, II, IV, I** This sequence ensures that the gene is properly introduced and accurately verified within the model organism.
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