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Figure 1. Schematic of SAGE method (Courtesy of sagenet.org)1, click to enlarge.

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.

Step-by-Step Guide to Serial Analysis of Gene Expression

DNA Purification kit

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:

PCR Kit

BigDye® Direct Cycle Sequencing Kit contains reagents to perform the PCR, PCR clean up and sequencing reactions.

Recommended Products:

DNA strands

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:

BigDye® Direct cycle sequencing kits

Sequencing Reaction Cleanup

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?

Cleanup Method Description & Application Product
Reagent-Based Sequencing Template CleanupScavenges all unincorporated BigDye® terminators. Stabilizes samples before analysis. Best for long and short fragment recovery.
  Instrument
 3730xl
3730xL
3730
3730
3500xl
3500xL
3500
3500
3130xl
3130xL
3130
3130
310
310
Number of Capillaries96482481641
Compatible Applications:
(S) Supported; (A) AB Demonstrated; (C) Customer Demonstrated; (N) Not Supported
Resequencing
SAGE™ Method CCCCCCA

Analyze the mitochondrial sequence.

Which Data Analysis Software Is Right for You?

Highlighted products for SAGE™:

Verti Thermal Cycler

3500 Series Genetic Analyzers

GeneMapper® Software