Application of cationic conjugated polymers in microarrays using label-free DNA targets.
A fluorescence-based microarray technique that does not require target DNA labeling is detailed. This 'label-free' approach utilizes a cationic, water-soluble conjugated polymer PFBT (poly[9,9'-bis(6''-(N,N,N-trimethylammonium)hexyl)fluorene-co-alt-4,7-(2,1,3-benzothiadiazole) dibromide]), and neutral PNA (peptide nucleic acid) hybridization probes. DNA hybridization to immobilized PNA spots results in a change in the net charge at that particular surface. Electrostatic interactions between the cationic polymer and negatively charged DNA bind the polymer to the hybrid DNA/PNA complex. By exciting the conjugated polymer at 488 nm on a commercial microarray scanner, the presence of the target is directly indicated by the fluorescence emission of the polymer. This feature eliminates the necessity of target labeling required in traditional microarray protocols. There are five steps involved in the procedure before scanning or imaging the array: (i) slide hydration, (ii) target hybridization, (iii) post-hybridization washing, (iv) polymer application and (v) polymer washing. Each step takes 20 min to 1 h. The overall protocol requires approximately 2-3 h.
Sun
ChengJun
C
Sirigen Inc., Santa Barbara, California 93111, USA.
Gaylord
Brent S
BS
Hong
Janice W
JW
Liu
Bin
B
Bazan
Guillermo C
GC
D052160:Benzothiazoles / Q000737:chemistry*; D004247:DNA / Q000032:analysis* / Q000737:chemistry; D015341:Nucleic Acid Probes / Q000737:chemistry; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D020135:Peptide Nucleic Acids / Q000737:chemistry; D000644:Quaternary Ammonium Compounds / Q000737:chemistry*
D052160:Benzothiazoles; D015341:Nucleic Acid Probes; D020135:Peptide Nucleic Acids; D000644:Quaternary Ammonium Compounds; C516930:poly((9,9-bis(6'-N,N,N-trimethylammoniumbromide)hexyl)fluorene-co-alt-4,7-(2,1,3-benzothiadiazole)); D004247:DNA