Dahlquist:DNA Microarray Protocol: Difference between revisions
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** [http://www.ambion.com/techlib/prot/fm_1753.pdf Ambion Protocol (PDF)]. Protocol packaged with kit. | ** [http://www.ambion.com/techlib/prot/fm_1753.pdf Ambion Protocol (PDF)]. Protocol packaged with kit. | ||
* CyDye Post-Labeling Reactive Dye Pack (GE Healthcare Life Sciences Catalog <nowiki>#</nowiki>RPN5661) | * CyDye Post-Labeling Reactive Dye Pack (GE Healthcare Life Sciences Catalog <nowiki>#</nowiki>RPN5661) | ||
** 12 X Cy3 + 12 X Cy5 | ** 12 X Cy3 + 12 X Cy5 per box | ||
** [http://www1.gelifesciences.com/aptrix/upp01077.nsf/Content/Products?OpenDocument&parentid=25800563&moduleid=164924 GE Healthcare Product Web Page]. Distributed through [http://www.vwrsp.com/catalog/product/index.cgi?catalog_number=95040-134&inE=1&highlight=95040-134&reference_type=1&partnumber=RPN5661&sim_code=1.0 VWR]. | ** [http://www1.gelifesciences.com/aptrix/upp01077.nsf/Content/Products?OpenDocument&parentid=25800563&moduleid=164924 GE Healthcare Product Web Page]. Distributed through [http://www.vwrsp.com/catalog/product/index.cgi?catalog_number=95040-134&inE=1&highlight=95040-134&reference_type=1&partnumber=RPN5661&sim_code=1.0 VWR]. | ||
** Use protocol for Amino Allyl MessageAmp™ II aRNA Amplification Kit. | ** Use protocol for Amino Allyl MessageAmp™ II aRNA Amplification Kit. | ||
Revision as of 16:41, 15 August 2008
Overview
This protocol describes all the steps necessary to perform aRNA synthesis, labeling, hybridization, and scanning of DNA microarrays for Saccharomyces cerevisiae, beginning with total RNA.
Materials
- Amino Allyl MessageAmp™ II aRNA Amplification Kit (Applied Biosystems Catalog #AM1753)
- 20 reactions per kit
- Applied Biosystems Catalog Web Page. Originally an Ambion product. Ambion was acquired by Applied Biosystems and the kit can only be purchased through them.
- Ambion Product Web Page. Contains useful links.
- Ambion Protocol (PDF). Protocol packaged with kit.
- CyDye Post-Labeling Reactive Dye Pack (GE Healthcare Life Sciences Catalog #RPN5661)
- 12 X Cy3 + 12 X Cy5 per box
- GE Healthcare Product Web Page. Distributed through VWR.
- Use protocol for Amino Allyl MessageAmp™ II aRNA Amplification Kit.
Procedure
- Step 1
- Step 2
- Step 2 has some additional information that goes with it. i.e. Keep at 4°C.
- Step 3
- Step 3 has multiple sub-steps within it.
- Enumerate each of those.
Notes
Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!
- List troubleshooting tips here.
- You can also link to FAQs/tips provided by other sources such as the manufacturer or other websites.
- Anecdotal observations that might be of use to others can also be posted here.
Please sign your name to your note by adding '''*~~~~''': to the beginning of your tip.
References
Relevant papers and books
- Goldbeter A and Koshland DE Jr. An amplified sensitivity arising from covalent modification in biological systems. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6840-4. DOI:10.1073/pnas.78.11.6840 |
- JACOB F and MONOD J. Genetic regulatory mechanisms in the synthesis of proteins. J Mol Biol. 1961 Jun;3:318-56. DOI:10.1016/s0022-2836(61)80072-7 |
- ISBN:0879697164
Contact
- Who has experience with this protocol?
or instead, discuss this protocol.
