Brown Lab Protocols

NEW protocols - June 2007

• Post-Processing of Oligo Arrays on Corning Ultra GAPS slides

• Post-Processing of Oligo Arrays on PolyLysine coated slides

• Ambion MessageAmp II Amino Allyl and Cy Dye Coupling for Oligo Arrays (PDF Version) (updated 05/18/2006)

• Preparation of Fluorescent DNA Probe from Total mRNA - Direct label of Total RNA for Oligo Arrays (PDF version) (updated 05/18/2006)

• Array Hybridization and Washing for Oligo Arrays (PDF version)

• Reverse Transcription (RT) and Amino-Allyl coupling of cDNA for (total) RNA sample input

Cy5 destruction has been reported after 10-30 seconds exposure to ozone levels above 5-10 ppb, especially during the array washing, drying and scanning steps. We have analyzed this problem and developed several solutions you can download here (updated June 2007):

• Details on ozone destruction of Cy5 and plans for building excellent ozoone-free enclosurese.

• A Good Cheap Kludge - how to build an ozone-free room.

• How to dry microarray slides with an ozone-free room and small centrifuge.

DNA Preparation

• Preparation of DNA Samples (updated 12/3/1999)

Slide Preparation

• Poly-l-Lysine coating of slides (updated 10/6/1999)