1. cDNA synthesis and microarray hybridization
Mod2 Day7
Tuesday Oct 28th

2. Notes from FNT About Figures: Use every opportunity to be informative!
Label strain names with relevant genotype
Tell a little about what you did to get generate the data

3. Notes from FNT
About calculation: if you measured A260= using 1:100
then A260 of undiluted =
2 ug x

4. Microarray: cDNA synthesis, microarray hybrization
Double ug RNA?
Anneal capture sequence 1, ° 10’
Extend with RT, 42° 1.5 hrs
Destroy RNA NaOH, mix on array

5. Microarrays: tomorrow
Wash off unhybridized cDNA
Hybridize with Cy3, Cy5 fluors
Wash off uncaptured fluors
Scan
Next time you’ll get a data file with info for 44K spots!

6. While you wait… Get to know what a data set will look like then
Normalization--why and how we do it
Background subtraction--what it’s for, what if you get a neg #
Log2 transformations--what it’s for and how we do it