Differences between the fingerprints generated from total RNA and poly-A RNA using a modified procedure of cDNA-AFLP and silver staining.
A modified procedure of cDNA-amplified fragment length polymorphism (cDNA-AFLP) combined with silver staining is described which generates clear band patterns from both fungal total RNA and poly-A RNA. The silver staining procedure takes only 25 min. Some 42-55 transcript-derived fragments (TDFs) were yielded from the total RNA of eight fungal pathogens. In comparison with the fingerprints generated from Mycosphaerella graminicola total RNA, those from poly-A RNA showed eight TDFs with an increased intensity and also seven additional TDFs. It suggests that poly-A RNA is preferable as starting material for ideal cDNA-AFLP analysis.
Institute of Phytopathology, Christian-Albrechts-University Kiel, Hermann-Rodewald-Str. 9, 24118 Kiel, Germany. jguo@phytomed.uni-kiel.de
Jian-Rong
JR
Guo
F
F
Schnieder
J A
JA
Verreet
D020552:Comet Assay / Q000379:methods*; D016172:DNA Fingerprinting / Q000379:methods*; D018076:DNA, Complementary / Q000235:genetics*; D012150:Polymorphism, Restriction Fragment Length; D012331:RNA, Fungal / Q000032:analysis* / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D016622:Silver Staining / Q000379:methods*
D018076:DNA, Complementary; D012331:RNA, Fungal; D012333:RNA, Messenger