Identification of Crassostrea ariakensis and related oysters by multiplex species-specific PCR

Genetic markers are needed for rapid and reliable identification of oysters. In this study, we developed multiplex genus- and species-specific PCR markers for the identification of oysters from China. We used the mitochondrial cytochrome oxidase I (COI) and nuclear 28S ribosomal RNA genes for marker development. DNA sequences from different species were obtained from GenBank or by direct sequencing. Sequences were aligned, and genus- and species-specific nucleotides were identified. Primers were designed for genus/species-specific amplification to generate fragments of different sizes. A multiplex set of genus- and species-specific primers from the 28S gene was able to separate C. ariakensis and C. hongkongensis from other species and assign oysters to four genera. A set of species-specific COI primers provided positive identification of all five Crassostrea species from China, C. ariakensis, C. hongkongensis, C. angulata, C. gigas, and C. sikamea in a single PCR. The multiplex PCR assays do not require fluorescence-labeling or post-PCR enzyme digestion, providing a simple, fast and reliable method for the identification of oysters from China.

基因芯片PCR温度模糊自整定PID控制算法的研究

在核酸扩增反应仪中,基因芯片核酸扩增反应过程要求实现温度高精度快速跟踪控制,常规温控方案和算法难以实现。将模糊推理系统与常规PID控制方式相结合,采用模糊自整定PID控制算法实现了温度快速跟踪控制。实验结果表明:模糊自整定PID控制算法比常规PID算法具有更强的鲁棒性,能够克服控制对象热惯性参数时变性的影响,降低了输出温度最大超调量,提高了稳态精度。

硫酸盐还原菌类群分离培养和PCR分析鉴定

探讨分离培养和聚合酶链反应（PCR）在硫酸盐还原菌类群分析鉴定中的应用。［方法］采用分离培养和PCR 对贵州阿哈湖沉积物中硫酸盐还原菌类群进行分析，并对分离纯化的一株硫酸盐还原菌进行鉴定。 ［结果］ 贵州阿哈湖沉积物硫酸盐还原菌类群为脱硫肠菌属、脱硫叶菌属和脱硫球菌- 脱硫线菌- 脱硫八叠菌属；纯化菌株经PCR 扩增初步鉴定为脱硫叶菌属,菌株形态与鉴定结果相符。 ［结论］ 采用分离培养结合PCR，可以对硫酸盐还原菌类群进行分析和鉴定。