ABSTRACT
Contents Introduction .............................................................................................................. 53
Substrate Selection .......................................................................................... 53 Immobilization by Adsorption ........................................................................ 53 Covalent Attachment .......................................................................................54 Quenching, Capping, and Blocking ................................................................54 Hybridization .................................................................................................. 55
List of Protocols ....................................................................................................... 55 Protocol 3.1: Evaluation of Microarray Substrates for Nucleic Acid Analysis ....... 55
How It Works .................................................................................................. 55 Required Materials .......................................................................................... 56 Reagent Preparation ........................................................................................ 56 Step-by-Step Protocol ..................................................................................... 56 Key References ............................................................................................... 57 Troubleshooting Guide ................................................................................... 57
Note 1: Slide Denaturation ................................................................. 57 Note 2: Hybridization ......................................................................... 57
Protocol 3.2: Noncovalent Adsorption of DNA to Amino-Silane Supports ............ 57 How It Works .................................................................................................. 57 Required Materials .......................................................................................... 58 Reagent Preparation ........................................................................................ 58 Step-by-Step Protocol ..................................................................................... 58 Key Reference ................................................................................................. 59 Troubleshooting Guide ................................................................................... 59
Note 1: Handling Issues ...................................................................... 59 Note 2: Source Plates .......................................................................... 59 Note 3: Use of HEPA Filter ................................................................ 59 Note 4: Capping and Blocking Residual Amines ............................... 59
Note 5: Capping Step .......................................................................... 59 Note 6: Denaturation ........................................................................... 59
Protocol 3.3: Covalent Attachment ..........................................................................60 Protocol 3.3.1: Covalent Coupling of Amino or Other Modied
Oligonucleotides to Solid Supports Containing Epoxides (Oxiranes) ...................................................................................................60
