Blocking Polylysines on Homemade Microarrays.

Area of Science:

• Biotechnology
• Molecular Biology
• Surface Chemistry

Background:

• Polylysine-coated slides are used for homemade microarrays.
• Lysine residues create a positively charged surface, leading to nonspecific binding of nucleic acids and high background fluorescence during hybridization.

Purpose of the Study:

• To develop a simple protocol for blocking surface lysines on microarray slides.
• To reduce background fluorescence in homemade microarrays for improved hybridization.

Main Methods:

• Succinylating the ε-amino group of lysine residues using succinic anhydride.
• Rehydrating slides (if necessary), drying, performing the succinylation reaction, washing, and drying with ethanol.

Main Results:

• The succinylation protocol effectively blocks surface lysines not associated with oligonucleotide spots.
• Reduced background fluorescence is achieved, preparing slides for hybridization.

Conclusions:

• This succinylation method is a straightforward and effective way to minimize background noise in homemade microarrays.
• The protocol enhances the quality of microarray data by improving the signal-to-noise ratio.