Full-length normalization subtractive hybridization: a novel method for generating differentially expressed cDNAs.
Here we present a novel method termed full-length normalization subtractive hybridization (FNSH) for efficiently generating subtracted cDNA libraries with a high degree of productivity. This method has the ability to isolate full-length differentially expressed genes from target samples. Normalization and subtraction of FNSH are performed simultaneously with efficiency equal to or even higher than that of suppression subtractive hybridization. Using FNSH, we have isolated at least 40 unique cDNAs that are expressed in terminal ampullae but not in the ovaries of the prawn Macrobrachium rosenbergii from 120 randomly picked subtracted clones. Sequence analysis shows that 37 of the 40 cDNAs are full length.
Dai
Zhong-Min
ZM
Institute of Cell Biology and Genetics, College of Life Sciences, Zhejiang University, Zijingang Campus, Hangzhou, Zhejiang, 310058, People's Republic of China.
Zhu
Xiao-Jing
XJ
Yang
Wei-Jun
WJ
D000818:Animals; D003001:Cloning, Molecular / Q000379:methods; D018076:DNA, Complementary / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D005260:Female; D008297:Male; D009693:Nucleic Acid Hybridization / Q000379:methods*; D009928:Organ Specificity; D010053:Ovary / Q000378:metabolism; D033541:Palaemonidae / Q000235:genetics; D013382:Subtraction Technique; D014649:Vas Deferens / Q000378:metabolism
D018076:DNA, Complementary