MAKING AN ARRAY

 

(Starting with a gold slide)

 

1. Soak in 1 mM MUAM for at least 2 hr.

 

 

2. Protect with Fmoc

 

 

a) dissolve the Fmoc in the DMSO, vortex, then add the TEA

b) cover surface of slide with this solution for 1 hour.

c) rinse with water

d) soak briefly in DMSO to remove excess Fmoc

 

3. Photopattern

 

 

a) place slide in holder, gold side up

b) place pattern on slide, copper side down

c) photopattern for 1 hour at 400 Watts

d) rinse with EtOH

 

4. Soak in 1mM MUAM for 2 hour

 

5. Spot on SSMCC

 

 

a) let sit 30 min.

b) rinse with water, dry

 

6. Spot on thiol-DNA

 

 

a)     let sit in humid environment for at least 2 hours; can let sit overnight

b)    rinse with water

 

7. Deprotect

 

a) submerge slide in 1M TAEA [tris (2-aminoethyl) amine] for 15 min.

b) rinse with water

 

8. "PEG"ylate

 

a) cover surface with 4mM PEG-NHS for 30 - 60 min

b) rinse with water

 

9. Store sample in phosphate buffer (20mM, pH 7.4) until ready to do imaging experiment

 

Copyright 1999 Corn Group

 

REFERENCE: J.M. Brockman et al. J. Am. Chem. Soc., 1999, 121, 8044-8051.