赛默飞世尔Thermo Fisher BECKMAN 384

详细BAC DNA Isolation

clone in the Floor Shaker for 18 hours at 37 degrees C shaking at 350 rpm. 2.For BAC or Fosmid isolation for end sequencing: Using the Hydra96, inoculate 2 μl of the BACs or Fosmids from their glycerol plate stocks into either a 96 or 384

详细BACDNAIsolation

rpm. 2.For BAC or Fosmid isolation for end sequencing: Using the Hydra96, inoculate 2 μl of the BACs or Fosmids from their glycerol plate stocks into either a 96 or 384 well shallow plate. Incubate at 37 deg C overnight

基因组高通量测序

建立在双脱氧测序法(Sanger等，1977)的基础上。为了从每一克隆插入片段两端成对地进行测序，每一个质粒模板DNA板应配备两个384孔循环测序反应板。测序反应采用Big Dye Terminator chemistry version 3．1(AppliedBiosystems)和标准M13或常用正向引物和反向引物。测序反应通过BiomekFX(Beckman)移液操作工作站建立。机械臂负责等分模板试样，起与反应液混合的作用，反应液含有双脱氧核苷酸、荧光标记的核苷酸、TaqDNA聚合酶、序列