cDNA synthesis and microarray hybridization Mod2 Day7 Tuesday Oct 28th

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

Notes from FNT About calculation: if you measured A260= 0.429 using 1:100 then A260 of undiluted = 2 ug x

Microarray: cDNA synthesis, microarray hybrization Double ug RNA? Anneal capture sequence 1, 2 80° 10’ Extend with RT, 42° 1.5 hrs Destroy RNA NaOH, mix on array

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!

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