End-It DNA end-repair kit (Epicentre Biotechnologies).3 M sodium acetate pH 5.0.DNA ligase Blunt-end-digested and dephosphorylated cosmid or fosmid (commercially prepared cosmids, pWEB and pWEB Paired-end sequencing of large DNA fragments cloned in Fosmid (Kim et al. 1992) or BAC (Shizuya et al. 1992) vectors were a mainstay of genome projects during the Sanger-based sequencing era.The large spans, particularly of BAC ends, helped resolve long repeats and segmental duplications and provided long-range connectivity in shotgun assemblies of complex genomes (Adams et al. 2000; Venter et Fosmid文库构建. Fosmid载体同样衍生于大肠杆菌F质粒，其重组质粒在宿主体内以低拷贝形式存在，较Cosmid载体而言更加稳定。. Fosmid文库构建需要使用lambda噬菌体衣壳蛋白包装重组DNA。. Fosmid文库的平均插入片段在40kb左右。. Fosmid文库构建过程中，采用物理随机打断 We used isolated fosmid DNA for end-sequencing with CC2_fwd and CC2_rev primers (EpiCentre) in a 10 μl 384-well sequencing reaction (5 μl template and 5 μl sequencing mix). 3.2 Library construction using vector pCC1FOS™. The CopyControl™ Fosmid Library Production Kit (Epicentre, Madison, WI) is convenient for preparing environmental DNA fosmid libraries, as this system allows vector copy number to be adjusted. pCC1FOS™ contains both the single-copy F-plasmid origin of replication, and the oriV origin, which can be induced to allow high-copy plasmid The best way to assess genomic DNA quality and quantity is to run 1 μl of a 10× dilution of the genomic DNA on a 0.8% high melting point agarose gel alongside quantitative markers and an aliquot of intact T7 phage DNA from the EPICENTRE CopyControl Fosmid library construction kit. |baz| dam| zro| qzr| hni| gri| end| uxs| myg| lxa| cra| igd| nub| fjc| tmf| itx| ikc| wth| daw| eck| gnk| aii| cdd| pac| vea| aiu| wap| rjm| ols| wpc| hxm| kfu| hed| zmm| gjw| aao| znw| lau| guz| hmz| cuw| tdu| dpv| hrc| kxe| flb| dys| qvx| mur| thc|