Analysis of small RNA populations using hybridization to DNA tiling arrays

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 631; p. 75
• Main Authors: Boccara, Martine, Sarazin, Alexis, Billoud, Bernard, Bulski, Agnes, Chapell, Louise, Baulcombe, David, Colot, Vincent
• Format: Journal Article
• Language: English
• Published: United States 01.01.2010
• Subjects: Arabidopsis - chemistry; Chromosomes, Plant - genetics; DNA, Complementary; DNA, Plant; Nucleic Acid Hybridization - methods; Oligonucleotide Array Sequence Analysis; RNA, Plant - analysis; RNA, Untranslated - analysis; Stress, Physiological - genetics
• ISSN: 1940-6029
• DOI: 10.1007/978-1-60761-646-7_8

Small RNA (sRNA) populations extracted from Arabidopsis plants submitted or not to biotic stress, were reverse-transcribed into cDNAs, and these were subsequently hybridized after labelling to a custom-made DNA tiling array covering Arabidopsis chromosome 4. We first designed a control experiment with eight cDNA clones corresponding to sequences located on chromosome 4 and obtained robust and specific hybridization signals. Furthermore, hybridization signals along chromosome 4 were in good agreement with sRNA abundance as previously determined by Massive Parallel Sequence Signature (MPSS) in the case of untreated plants, but differed substantially after stress treatment. These results demonstrate the utility of hybridization to DNA tiling arrays to detect major changes in small RNA populations.