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【讨论】生物纳米颗粒与肿瘤的基因治疗

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【公告】Cytonacx 和癌症的基因治疗

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  • mpark
    mpark  回复了帖子 请问这里lnc编码能力 构建质粒怎么证明(如图)? 46分钟前
    The three expression constructs are same except thet the start ATG is placed in three different coding position or in three different open reading frame (ORF) to test translation of potential peptides. The testing construct are introduced into cells by transfection, and the cellular protein extracts are prepared and probed with anti-myc antibody in
  • mpark
    mpark  回复了帖子 咨询质粒转化步骤源材料质粒选择问题 56分钟前
    You can try both, but run minipreps to screen and confirm the plasmid.
  • mpark
    mpark  回复了帖子 小鼠有表型,原代细胞检测不到该基因的变化 1天前
    我们用小鼠提取原代细胞,并体外培养3-5天后,用q-pcr检测该基因mRNA表达,发现没变化, for a gene knockout mouse, try to run Southern blot or conventional PCR to verify the composits at the gene loci.
  • mpark
    mpark  回复了帖子 求助基因作图方法!!! 1天前
    You can try to make this picture by running  MicroSoft Office PowerPoint program.
  • mpark
    mpark  回复了帖子 芯片处理 1天前
    Use the link:http://dxys.com/psLHVl lumi package provides an integrated solution for the    Illumina microarray data analysis. It includes functions of    Illumina BeadStudio (GenomeStudio) data input, quality control,     BeadArray-specific variance stabilization, normalization an
  • mpark
    mpark  回复了帖子 哪个大神给解释一下CRE consensus是啥意思? 1天前
    CRE consensus is a track of DNA nucleotide sequences which contain core or essential nucleotides specifically for associating with CRE protein.
  • mpark
    mpark  回复了帖子 质粒双酶切只有一个条带怎么办? 4天前
    Try to have the construct DNA sequenced, make sure that the two enzume sites are correct and located in right positions, and the insert DNA size is right. If cut 1 ug DNA, the cut off 0.3 kb fragment is hard to see on the gel, try cut 10 ug DNA in a final volumes of 200-500 ul. Run 2-2.5 % agarose gel  (using low meltng agarose) to purify the
  • mpark
    mpark  回复了帖子 求助:双荧光素酶突变质粒荧光值要高太多 6天前
    为什么转染结合位点突变的质粒要比野生型质粒的值高两倍多: try to find information about transcription regulation regarding to this DNA fragment mutated at the transcription factor or factors binding site, if gel shift assay or DNase I footprinting shows stronger association of transcription factor or factors with the DNA mutated at DNA binding sites than at the DNA in wild typ
  • mpark
    mpark  回复了帖子 求助!!!!用T7E1死活切不开。 6天前
    You need to isolate Cresrp Cas 9 /sgRNA transfected cells or cell lines before running T7E1 cleavage assay. The other issue is that PCR amplification of DNAv from sgRNA transfected cells or cell lines would let you run T7E1 easily.Please download the attached file for details.
  • mpark
    mpark  回复了帖子 关于GFP发光的问题 10天前
    To make GFP green cells, the transfection efficiency would be above 85% or even high. After transfection, you may perform cell sorting to enrich GFP expression green cells, or you may run cell colony selection to isolate cell clones with high levels of GFP expression.
  • mpark
    mpark  回复了帖子 Chip求助 11天前
    如果不去背景的话, 我的结果着丝粒处富集的蛋白和染色体臂富集差不多甚至要少一些: you have said that protein A associates with DNA nonspecifically, it indicates that delta t values of no-antibody IPs should be quite same whether DNA at the arms or at CENs. Your results seem to be discrepancy from this assumption.
  • mpark
    mpark  回复了帖子 Chip求助 11天前
    CHIP without adding antibody should not have any crosslinked chromatin DNA amplified, therefore the delta t will be zero. In this way, how the background would be eliminated?
  • mpark
    mpark  回复了帖子 关于GFP发光的问题 11天前
    Try to run good transfection for GFP expression.
  • mpark
    mpark  回复了帖子 关于GFP发光的问题 12天前
    是不是曝光时间太长: GFP fluorescence excitation spectrum is 488 nm, and its emission 509 nm. As long as GFP protein is in good condition, the blue fluorescence emission will be detected in proportion to amounts of GFP protein.  
  • mpark
    mpark  回复了帖子 Chip求助 12天前
    如果去背景的话着丝粒处富集的蛋白确实比染色体臂富集的多: can you try to explain more about Background? What does background mean, non specific PCR band or bands? 
  • mpark
    mpark  回复了帖子 求助: EMSA的阻滞带位置不太正常 12天前
    That is a specific shift formed by the 400 DNA probe and the tested protein. Try to run gel shift using increased amounts of tested protein and constant amounts of DNA probe to optimize the association of protein and the DNA.If it is possible, try to break down the 400 bp DNA probe into 2 or 3 smaller pieces and test each shortened DNA in gel shift
  • mpark
    mpark  回复了帖子 蛋白和dna结合 13天前
    Please describe what it the fus启动子的基因. To confirm the protein and DNA association, run competition tests in gel shift assays.
  • mpark
    mpark  回复了帖子 求助: EMSA的阻滞带位置不太正常 13天前
    Please run competition tests using cold 400 bp DNA fragments in at least 50 folds molar ratio to the probe DNA.
  • mpark
    mpark  回复了帖子 羊穿结果回报提示提示13号染色体上有13.8Mb基因为纯合片段,临床意义不明确,求各位老师指导 13天前
    The results presented are in poor quality and  could not be reviewed for details. Try to upload original data in PDF files
  • mpark
    mpark  回复了帖子 羊穿结果回报提示提示13号染色体上有13.8Mb基因为纯合片段,临床意义不明确,求各位老师指导 13天前
    Try to ask the Lab to repeat chip hybridization and whole genome sequencing using samples taken from differewnt time.

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