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Ch 8.Immunohistoch / immunology
Ch 10.GC/MS, NMR and Proteomics
Molecular Biology / PCR / Variants of PCR / RACE PCR
Background: Rapid amplification of cDNA ends (RACE) is a polymerase
chain reaction-
based technique which facilitates the cloning of full-length cDNA
sequences when only a partial cDNA sequence is available.
Traditionally,
cDNA sequence is obtained from clones isolated from plasmid or
phage libraries. Frequently these clones lack sequences
corresponding to
the 5' ends of the mRNA transcripts. The missing sequence information
is typically sought by repeatedly screening the
cDNA library in an effort
to obtain clones that extended further towards the 5' end of the message.
The nature of the enzymatic reactions
employed to produce cDNA
libraries limits the probability of retrieving extreme 5' sequence even
from libraries that are very high quality.
RACE-PCR
facilitates the isolation of 5 and 3 end sequences from cDNA.
Human Molecular Genetics 2 conveys the feel of fast-moving research while
providing a description in some depth of the techniques and data that are helping ...
www.ncbi.nlm.nih.gov/books/bv.fcgi?rid=hmg.figgrp.2574
5' RACE PCR Protocol 1. First strand cDNA synthesis: - Reaction set up: cDNA synthesis buffer 4 ul dNTP mixture 2 ul gene
specific primer 1 1 ul ...
biowww.net/detail-126.html
Detailed protocol on 3' RACE PCR - rapid amplification of 3'
cDNA end. ... 3' RACE PCR Protocol: 1. First strand cDNA synthesis: ...
biowww.net/detail-329.html
SMART RACE cDNA Amplification Protocol-at-a-Glance
Protocol-at-a-Glance.
2. For 5'-RACE and 3'-RACE: prepare PCR reactions as shown in Tables III & IV. Add the components in the order shown in PCR tubes.
...
www.clontech.com/images/pt/PT3269-2.pdf
5' RACE Protocol For Generation of Seq. Tags.
Apply all 79 ul of1st PCR product to
center of packed wells. ... Sequence template per 5'RACE sequencing protocol with sequencing oligo 305. ...
baygenomics.ucsf.edu/protocols/comp1/RACE_generation.pdf
Rapid Amplification of cDNA 3' Ends (3'- RACE)
( Simplified protocol based on Clontech¡¯s SMART- RACE ). 6. Distribute 5 ¦Ìl
of diluted first PCR product in the PCR tubes, 8. strips or 96 plate on ice. ...
www.cmhd.ca/protocols/genetrap_pdf/3'RACE.pdf
A
lock-docking oligo(dT) primer for 5' and 3' RACE PCR.
stretch of As, yielding a discrete PCR. product. We are planning to use the lock-dock-.
ing primer in the 5'-RACE protocol (1) to ...
www.genome.org/cgi/reprint/2/2/144.pdf
First Choice RACE-Ready cDNA Protocol
The
3' RACE protocol. describes nested PCR, however 3' RACE reactions may produce signif- ..... your PCR protocol. Figure 3. 3' RACE Control.
3' Adapter ...
http://www.ambion.com/techlib/prot/fm_3200.pdf
Nature Protocols: 3¡ä End cDNA amplification using classic RACE
...
amplification of cDNA ends (RACE) PCR as long as part of the mRNA sequence is known; ... cDNA ends can be obtained in 1¨C3 days using
this protocol. ...
www.natureprotocols.com/2007/01/11/3_end_cdna_amplification_using.php
High throughput Rapid Amplification of
cDNA Ends (RACE) using cDNA Library as Template.
Follow standard protocol for plasmid prep in microtitterplates. PCR for RACE-clones:.
Use standard protocol for PCR, but use 1 ul plasmidprep as template ...
www.bio.aau.dk/download/biotechnology/kln/RACE-procedures.doc
Help for 3'-RACE PCR. PROBLEM: We have prolem to amplify the 3'-end of our .... I intend to try the following: a) Modified
an inverse PCR protocol by ...
http://wheat.pw.usda.gov/~lazo/methods/lazo/met11.html