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epicentre HTFP061 pCC2 Forward Sequencing Primer

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  • 品  牌:
  • epicentre HTFP061
  • 主要规格:
  • 1 nmole
  • 用  途:
  • 科研
    • epicentre HTFP061 pCC2 Forward Sequencing Primer 产品简介:北京中北林格供应epicentre HTFP061 pCC2 Forward Sequencing Primer,中北林格是epicentre总代理,提供epicentre产品货号报价查询。其他包括lucigen总代理,cellscript总代理,Biosearch technologies总代理,epicentre总代理,illumina总代理,ICLLAB总代理,ImmunoReagents总代理。

      供应商:中北林格

      产地:us

      发货地:北京

      10 9 pfu / µg DNA)包装,并包被在试剂盒中提供的TransforMax™EPI300™ 大肠杆菌(图3)。 The kit uses a strategy of cloning blunt-ended DNA fragments generated by random shearing of the DNA, to produce more complete and unbiased genomic libraries than can be obtained by partial restriction endonuclease digests. Genomic DNA is first sheared into approximately 40-kb fragments. The sheared DNA is end-repaired to generate blunt, 5´-phosphorylated ends and then size-selected by and recovered from a low-melting-point agarose gel. Finally, the size-selected DNA is ligated into the cloning-ready CopyControl pCC1FOS or pCC2FOS Vector, packaged using ultra-high efficiency MaxPlax™ Lambda Packaging Extracts (>109 pfu/µg DNA), included in the kit, and plated on the supplied TransforMax™ EPI300™ E. coli (Fig. 3). 好处说明 可提供CopyControl的CopyControl pCC1FOS和pCC2FOS载体:线性化,去磷酸化,纯化并准备连接。 CopyControl pCC1FOS and pCC2FOS Vectors are supplied Cloning-Ready: linearized, dephosphorylated, purified, and ready for ligation. 无需部分限制核酸内切酶消化或脉冲场凝胶电泳即可制备用于克隆的基因组DNA。 No need for partial restriction endonuclease digests or pulse field gel electrophoresis to prepare the genomic DNA for cloning. 使用pCC2FOS载体最大化高通量末端序列结果(图4)。 Maximize high-throughput end-sequence results using the pCC2FOS Vector (Fig. 4). 克隆可以从单个拷贝诱导到每个细胞多达50个拷贝(图5)。安全地获得更高的DNA产量,同时保持单拷贝数克隆的稳定性。 Clones can be induced from single copy up to 50 copies per cell (Fig. 5). Safely obtain higher DNA yields while maintaining the stability of single-copy-number clones. 高效的λ包装消除了背景和误报。 High-efficiency lambda packaging eliminates background and false positives. 比BAC克隆更快更容易。 Faster and easier than BAC cloning. epicentre HTFP061 pCC2 Forward Sequencing Primer 图例说明 图1. CopyControl™矢量地图。用于CopyControl Fosmid库生产的CopyControl pCC1FOS™和pCC2FOS™载体在Eco 72 I(平端)位点线性提供,然后脱磷酸化。该载体已准备好克隆约40 kb的末端修复(平末端)基因组DNA。 Figure 1. CopyControl™ Vector map. The CopyControl pCC1FOS™ and pCC2FOS™ Vectors for CopyControl Fosmid library production are supplied linearized at the Eco72 I (blunt) site and then dephosphorylated. The vector is ready for cloning end-repaired (blunt-end) genomic DNA of approximately 40 kb. 图2. CopyControl™pCC2FOS™矢量引物盒。该载体与pCC1FOS载体的区别在于新引物盒的工程化,该引物盒消除了浪费的源自载体的测序读数,并最大程度地减少了在大肠杆菌基因组上引发的可能性。 Figure 2. The CopyControl™ pCC2FOS™ Vector primer cassette. The vector differs from the pCC1FOS Vector by the engineering of a new primer cassette that eliminates wasteful vector-derived sequencing reads and minimizes the potential for priming on the E. coli genome. 图3(单击放大)。使用CopyControl™Fosmid库生产工具包准备fosmid库的过程概述。一旦准备好文库,就可以使用EPICENTRE的DirectLysis Fosmid96试剂盒或FosmidMAX™DNA纯化试剂盒,以小体积培养单个克隆,并诱导成多拷贝数,从而获得高产率的高纯度DNA,用于指纹,测序等。 Figure 3 (click to enlarge). Overview of the process for preparing a fosmid library using the CopyControl™ Fosmid Library Production Kits. Once the library has been prepared, individual clones can be cultured in small volume and induced to multiple-copy number for high yields of high-purity DNA for fingerprinting, sequencing, etc., using EPICENTRE's DirectLysis Fosmid96 kit or FosmidMAX™ DNA Purification Kit. 图4.用pCC2FOS™正向引物在pCC2FOS™克隆上以1 / 48x BigDye™稀释度获得的典型测序结果。用pCC2FOS反向引物获得了相似的结果(数据未显示)。 Figure 4. Typical sequencing results obtained with the pCC2FOS™ Forward Primer on a pCC2FOS™ clone at 1/48x BigDye™ dilution. Similar results were obtained with the pCC2FOS Reverse Primer (data not shown). 图5.每个细胞最多可复制50个拷贝的CopyControl™Fosmid克隆,以大大提高DNA产量。 从未诱导的(–)和诱导的(+)CopyControl克隆中分离出的fosmid DNA的Hin d III消化物。消化液占样品总体积的三分之一(8 µl),并通过琼脂糖凝胶电泳进行分析。M线,千磅梯子。 Figure 5. CopyControl™ Fosmid clones can be induced up to 50 copies per cell to greatly increase DNA yield. Hind III digests of fosmid DNA isolated from uninduced (–) and induced (+) CopyControl clones. Digests contained one-third (8 µl) of the total sample volume and were analyzed by agarose gel electrophoresis. Lane M, Kilobase ladder. 对此产品有兴趣,可联系中北林格获取更多详细信息。

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