Construction and characterization of cDNA library from water-stressed plantlets regenerated in vitro of Populus hopeiensis
2005
In order to isolate and clone water-stress-responsive genes, total RNA was extracted from water-stressed plantlets regenerated in vitro of Populus hopeiensis using a QIAGEN RNeasy Plant Mini Kit. CDNA, synthesized by LD-PCR with the SMART cDNA Library Construction Kit, was in vitro packaged into a phage λTriplEx2 vector. The resulting primary library and amplified library have a titer of 1.68 × 106 and 1.69× 109 pfu·mL−1 respectively. The combination ratio reached 98.8% and the average size of inserts was about 800 bp. In addition, the percentage of inserted fragments (>400 bp) was approximately 90%. The results indicate that a cDNA library has been successfully constructed.
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