Construction and Indentification of Full-length cDNA Library from Young Pod of Soybean Jinong 18

Construction of full-length cDNA library from soybean young pod could lay foundation for cloning yield related genes.The total RNA from young pod of soybean Jinong 18 was separated and then transcribed into single-strand cDNA,which was synthesized into double-strand cDNAs by LD-PCR.Double-strand cDNAs was digested by Proteinase K and Sfil,and fractionated by CHROMA SPIN-400 columns.The ds cDNAs longer than 500 bp were collected and ligated to λTripIE × 2,and then packaging reaction for recombinant bacteriophages was performed.The titer of the primary cDNA library was 1.5 × 106pfu.mL-1,and the recombinant rate was 92%.The titer of amplified library was 2.6 × 108 pfu.mL-1and the insert size was more than 500 bp.Results showed that library titer and recombination rate met requirements of cDNA library,which would facilitate the functional genomics research of soybean.