Abstract：To investigate the stability of reference molecules with different insertion segments, there standard plasmids were developed with different insert segments: 186 bp (Cry1Ab), 273 bp(Cry1Ab and SPS) and 880 bp(Cry1Ab, SPS, fsACP, 35S, NOS and NPTII). The plasmids were stored for different time (1, 3, 7, 14, 21, 28 days or 3, 6, 12 months) under different temperature (-70, -20 and 4℃). The stability of Cry1Ab of 186, 273 and 880 bp were compared. The results showed, the Cq(quantification cycle) values of Cry1Ab in 186 and 273 bp were stable in 6 months, but increased rapidly and significantly differed (P<0.05) under -70℃ and 4℃ storing for 12 month. The Cq values of Cry1Ab in 880 bp were increased and significantly differed (P<0.05) at the 6 month. The stability of SPS in 273 bp and SPS, fsACP, 35S, NOS, NPTII in 880 bp were also studied, and we found that the Cq values of SPS in 273 bp were stable in 6 months, and the Cq values of the other five segments in 880 could maintain stability only in 3 months. It was showed that plasmid with shorter insert segments were more stable than that with the longer one, and the stability varied with different insertion. This study provides a scientific basis of the stability of reference molecules.
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