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Serial Analysis of Gene Expression (SAGE) by Sanger Sequencing
The SAGE™ technique is one of the more comprehensive methods available for detailed analysis of vast number of cellular transcripts. Initially developed to study differential gene expression in colon cancer samples, SAGE™ analysis compares quantitative expression data for multiple genes in a given specimen to create accurate relative expression profiles.
The ability to count many thousands of genes means that you can detect genes expressed at very low levels in a high-throughput manner. The three-step SAGE™ process allows for simultaneous analysis of sequences derived from various cell populations or tissues.
Figure 1. Schematic of SAGE method (Courtesy of sagenet.org)1, click to enlarge.
Step-by-Step Guide to Serial Analysis of Gene Expression
 DNA extraction is a critical first step in the experimental workflow of DNA Sequencing and Fragment analysis. The overall quality, accuracy and length of the DNA sequence read can be significantly affected by characteristics of the sample itself, and the method chosen for nucleic acid extraction. Ideal methods will vary depending on the source or tissue type, how it was obtained from its source, and how the sample was handled or stored prior to extraction. Recommended Products: |
 BigDye® Direct Cycle Sequencing Kit contains reagents to perform the PCR, PCR clean up and sequencing reactions. Recommended Products: |
 BigDye® Direct Cycle Sequencing Kit contains reagents to perform the PCR, PCR clean up and sequencing reactions. PCR Clean up and sequencing reaction are perform in a single step. Recommended Products: |
 After the sequencing reaction, it is important to remove unincorporated dye terminators and salts that may compete for capillary electrophoretic injection. Unincorporated terminators can co-migrate with the sequencing template, resulting in basecalling er Recommended Products:Which Sample Cleanup Product Is Right for You?
|
 Analyze the mitochondrial sequence. Which Data Analysis Software Is Right for You?
|
DNA extraction is a critical first step in the experimental workflow of DNA Sequencing and Fragment analysis. The overall quality, accuracy and length of the DNA sequence read can be significantly affected by characteristics of the sample itself, and the method chosen for nucleic acid extraction. Ideal methods will vary depending on the source or tissue type, how it was obtained from its source, and how the sample was handled or stored prior to extraction. Recommended Products: |
BigDye® Direct Cycle Sequencing Kit contains reagents to perform the PCR, PCR clean up and sequencing reactions. Recommended Products: |
BigDye® Direct Cycle Sequencing Kit contains reagents to perform the PCR, PCR clean up and sequencing reactions. PCR Clean up and sequencing reaction are perform in a single step. Recommended Products: |
After the sequencing reaction, it is important to remove unincorporated dye terminators and salts that may compete for capillary electrophoretic injection. Unincorporated terminators can co-migrate with the sequencing template, resulting in basecalling er Recommended Products:Which Sample Cleanup Product Is Right for You?
|
Analyze the mitochondrial sequence. Which Data Analysis Software Is Right for You?
|
- Publications: Serial Analysis of Gene Expression in Differentiated Cultures of Human Epidermal Keratinocytes.
Journal of Investigative Dermatology (2001)
116, 12–22; doi:10.1046/j.1523-1747.2001.00218.x
Step-by-Step Guide to Serial Analysis of Gene Expression
DNA extraction is a critical first step in the experimental workflow of DNA Sequencing and Fragment analysis. The overall quality, accuracy and length of the DNA sequence read can be significantly affected by characteristics of the sample itself, and the method chosen for nucleic acid extraction. Ideal methods will vary depending on the source or tissue type, how it was obtained from its source, and how the sample was handled or stored prior to extraction. Recommended Products: |
BigDye® Direct Cycle Sequencing Kit contains reagents to perform the PCR, PCR clean up and sequencing reactions. Recommended Products: |
BigDye® Direct Cycle Sequencing Kit contains reagents to perform the PCR, PCR clean up and sequencing reactions. PCR Clean up and sequencing reaction are perform in a single step. Recommended Products: |
After the sequencing reaction, it is important to remove unincorporated dye terminators and salts that may compete for capillary electrophoretic injection. Unincorporated terminators can co-migrate with the sequencing template, resulting in basecalling er Recommended Products:Which Sample Cleanup Product Is Right for You?
|
Analyze the mitochondrial sequence. Which Data Analysis Software Is Right for You?
|
DNA extraction is a critical first step in the experimental workflow of DNA Sequencing and Fragment analysis. The overall quality, accuracy and length of the DNA sequence read can be significantly affected by characteristics of the sample itself, and the method chosen for nucleic acid extraction. Ideal methods will vary depending on the source or tissue type, how it was obtained from its source, and how the sample was handled or stored prior to extraction. Recommended Products: |
BigDye® Direct Cycle Sequencing Kit contains reagents to perform the PCR, PCR clean up and sequencing reactions. Recommended Products: |
BigDye® Direct Cycle Sequencing Kit contains reagents to perform the PCR, PCR clean up and sequencing reactions. PCR Clean up and sequencing reaction are perform in a single step. Recommended Products: |
After the sequencing reaction, it is important to remove unincorporated dye terminators and salts that may compete for capillary electrophoretic injection. Unincorporated terminators can co-migrate with the sequencing template, resulting in basecalling er Recommended Products:Which Sample Cleanup Product Is Right for You?
|
Analyze the mitochondrial sequence. Which Data Analysis Software Is Right for You?
|
- Publications: Serial Analysis of Gene Expression in Differentiated Cultures of Human Epidermal Keratinocytes.
Journal of Investigative Dermatology (2001)
116, 12–22; doi:10.1046/j.1523-1747.2001.00218.x
Highlighted products for SAGE™:
1. Johns Hopkins University SAGE Web site: http://www.sagenet.org.
Schematic of SAGE method.
April 2001.
For Research Use Only. Not for use in diagnostic procedures.