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The cDNAs were derived from reverse transcription of mRNA samples from seeds at different stages of germination and seedlings at early phase of growth under chilling stress (13oC/10oC). The mRNA pool was used as template for double stranded cDNA synthesis using the Stratagene Uni-Zap XR cDNA synthesis and library kit. A total of 150,000 phages were excised from the primary library as pBluescript phagemid clones. Enrichment of the primary excised library with chilling-induced transcripts was performed by hybridizing the primary excised library colony lifts with the PCR-select subtraction product, with cold germinated cDNA as tester and control temperature-germinated cDNA as driver.
Nucleotide
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