三倍体枇杷叶绿体DNA提取方法的优化

将三倍体枇杷叶片通过液氮研磨、差速离心与蔗糖密度梯度离心相结合的方法得到高纯度的叶绿体,在光学显微镜和荧光显微镜下分别观察到椭圆形颗粒状和自发红色荧光的叶绿体.提取的cpDNA经凝胶电泳检测,完整性好,纯度高,无降解.经紫外分光光度仪测定,所得cpDNA浓度为0.761ng/ul.

Optimization of the Methods for Extracting Triploid Locquat cpDNA

High purity chloroplast of triploid loquat was extracted by grinding leaves in liquid nitrogen,combining of differential centrifugation(400×g centrifugal 5 min,800×g centrifugal 6 min) and sucrose density gradient(52%,30%) centrifugation.Oval granular and spontaneous red fluorescence chloroplasts were observed under the optical microscope and the fluorescence microscope,respectively.Gel electrophoresis showed that the cpDNA thus extracted had good integrity and high purity and no degradation occurred.By UV spectrophotometry,the concentration of the cpDNA was 0.761 ng/μl.