Dahlquist:DNA Microarray Protocol: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
No edit summary |
|||
| Line 51: | Line 51: | ||
==References== | ==References== | ||
'''Relevant papers and books''' | '''Relevant papers and books''' | ||
If this protocol has papers or books associated with it, list those references here. See the [[OpenWetWare:Biblio]] page for more information. | |||
<!-- | |||
<biblio> | <biblio> | ||
#Goldbeter-PNAS-1981 pmid=6947258 | #Goldbeter-PNAS-1981 pmid=6947258 | ||
#Jacob-JMB-1961 pmid=13718526 | #Jacob-JMB-1961 pmid=13718526 | ||
#Ptashne-Genetic-Switch isbn=0879697164 | #Ptashne-Genetic-Switch isbn=0879697164 | ||
</biblio> | </biblio> --> | ||
==Contact== | ==Contact== | ||
Revision as of 23:53, 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.
NOTE: This protocol is still under construction!
Materials
Supplies & Reagents
- 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.
- TE Buffer
- Microcuvettes
- Nuclease-free water (non-DEPC treated)
- Fragmentation Reagents
- DIG Easy Hyb
- Sonicated salmon sperm DNA
- oligo dA
Equipment
- Dedicated set of RNase-free pipetmen
- Speedvac
- Water bath set to 50°C
- Water bath set to 37°C
- Heat block set to 70°C
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
If this protocol has papers or books associated with it, list those references here. See the OpenWetWare:Biblio page for more information.
Contact
- Who has experience with this protocol?
or instead, discuss this protocol.
