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GSL - New Applications
Serial
Analysis of Gene Expression
(An Alternative for Microarrays)
Serial Analysis of Gene Expression (SAGE)
is a sequence-based approach to the identification of differentially
expressed genes by comparative analysis. SAGE is a powerful tool, one of the
more comprehensive methods available for detailed analysis of large numbers
of cellular transcripts, leading to a profile of the expressed genes. SAGE
provides an accurate quantitative analysis of the relative levels of genes
expressed in a specimen. The ability to count many thousands of genes allows
the detection of genes that are expressed at very low levels in a
high-throughput manner. A three-step molecular approach, the SAGE process
allows simultaneous analysis of sequences derived from various cell
populations or tissues.
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SAGE is a
well-recognized method of gene-expression profiling. The information
gathered from SAGE analysis enables a clear view of the genes
associated with normal, developmental, and disease states.
Researchers require an efficient means of accessing the sequence
information contained in the concatamer of transcript tags.
Invitrogen Corporation has
released
I-SAGE, the first pre-assembled kit for performing SAGE
technology. This all-inclusive kit provides sufficent pre-tested
materials to generate up to six SAGE libraries for subsequent
sequencing.
For more detailed description
of SAGE, please visit:
www.sagenet.org
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Human Identification
Applications
Applied Biosystems has combined the
advantages of PCR-based testing and fluorescence detection into a highly
discriminating kit designed for human identification applications. The
AmpFLSTR® Profiler Plus™ PCR Amplification Kit amplifies nine
tetranucleotide short tandem repeat (STR) loci and the Amelogenin locus in a
single reaction tube. The STR loci amplified are D3S1358, D5S818, D7S820,
D8S1179, D13S317, D18S51, D21S11, FGA, and vWA. The Amelogenin locus is used
for gender identification because products of different lengths are
generated from the X and Y chromosomes.
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The AmpFLSTR®
Profiler Plus™ reagents and protocols have been developed to produce
the quality results necessary for human identification applications.
To increase the success of obtaining results in the presence of
enzyme inhibitors, bovine serum albumin (BSA) has been included in
the AmpFLSTR® PCR Reaction Mix.
Thermal cycler times and
temperatures have been developed to produce specific amplification
while providing the necessary degree of sensitivity. The recommended
range of input DNA is 1.0 to 2.5 ng. Furthermore, instrument
detection protocols have been provided to yield reproducible results
with excellent resolution and precision in sizing alleles. |
Amplified Fragment Length Polymorphism
AFLP® is a DNA fingerprinting
technique which detects DNA restriction fragments by means of PCR
amplification.
The AFLP® technology usually comprises
of the following steps:
The restriction of the DNA with two
restriction enzymes, preferably a hexa-cutter and a tetra-cutter;
The ligation of double-stranded (ds) adapters to the ends of the restriction
fragments;
The amplification of a subset of the restriction fragments using two primers
complementary to the adapter and restriction site sequences, and extended at
their 3' ends by "selective" nucleotides (see Figures 1 and 2);
Gel electrophoresis of the amplified restriction fragments on denaturing
polyacrylamide gels ("sequence gels");
The visualization of the DNA fingerprints by means of autoradiography,
phospho-imaging, or other methods. For more information about AFLP please
visit:
http://www.keygene.com/html/aflp.htm
Sequence
Analysis Tools
For the DNA motif analysis please
visit Wheaton College's
genomics research group
For searching public databases for
protein and nucleic acid sequences, pleasse visit
the San Diego Supercomputer Center's
Biology Workbench
For sequence manipulation, please
visit "Sequence Manipulation Suit"
For protein researchers, proteomics
tools can be found at "Expert Protein
Analysis System"
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