Screening of cDNA libraries on glass slide microarrays.
A quantitative screening method was developed to evaluate the quality of cDNA libraries constructed by suppression subtraction hybridization (SSH) or other enrichment techniques. The SSH technique was adapted to facilitate screening of the resultant library on a small number of glass slide microarrays. A simple data analysis pipeline named SSHscreen using "linear models for microarray data" (limma) functions in the R computing environment was developed to identify clones in the cDNA libraries that are significantly differentially expressed, and to determine if they were rare or abundant in the original treated sample. This approach facilitates the choice of clones from the cDNA library for further analysis, such as DNA sequencing, Northern blotting, RT-PCR, or detailed expression profiling using a custom cDNA microarray. Furthermore, this strategy is particularly useful for studies of nonmodel organisms for which there is little genome sequence information.
Berger
Dave K
DK
Department of Botany, Forestry and Agricultural Biotechnology Institute, University of Pretoria, South Africa.
Crampton
Bridget G
BG
Hein
Ingo
I
Vos
Wiesner
W
D003001:Cloning, Molecular; D020869:Gene Expression Profiling / Q000379:methods*; D015723:Gene Library; D005898:Glass / Q000737:chemistry*; D009693:Nucleic Acid Hybridization / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D031782:Pennisetum / Q000235:genetics* / Q000378:metabolism; D018515:Plant Leaves / Q000378:metabolism / Q000382:microbiology; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D018749:RNA, Plant / Q000235:genetics*; D013489:Suppression, Genetic / Q000235:genetics
D010940:Plant Proteins; D018749:RNA, Plant