• 我要登录|
  • 免费注册
    |
  • 我的丁香通
    • 企业机构:
    • 成为企业机构
    • 个人用户:
    • 个人中心
  • 移动端
    移动端
丁香通 logo丁香实验_LOGO
搜实验

    大家都在搜

      大家都在搜

        0 人通过求购买到了急需的产品
        免费发布求购
        发布求购
        点赞
        收藏
        wx-share
        分享

        Analysis of Gene Expression in Bone by Quantitative RT-PCR

        互联网

        433
        The isolation of undegraded RNA, free from inhibitors of reverse transcription-polymerase chain reaction (RT-PCR), is a major technical challenge in the analysis of gene expression in the skeleton. Bone is a mineralized tissue containing an abundant matrix, which makes RNA isolation difficult. On the positive side, however, frozen bone is quite brittle and can be ground to a powder, thus releasing the cell contents. Reno and colleagues (1 ) evaluated 12 different protocols for isolating total RNA from small amounts of rabbit ligament, cartilage, and tendon using phenol-guanidinium-based reagents. Like bone, these tissues contain few cells in an abundant organic matrix. Absence of ribosomal bands, nondetection of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA on Northern blotting, presence of DNA and/or protein contamination, and insoluble RNA pellets generated by these procedures led to the development of the Trispin method, based on a combination of two commercially available kits (2 ). In this procedure, the tissue is powdered in a stainless steel ball mill vessel that is cooled in liquid nitrogen. We have used a ball mill and modified Trispin method to extract high-quality, RT-PCR-ready RNA from bone samples.
        ad image
        提问
        扫一扫
        丁香实验小程序二维码
        实验小助手
        丁香实验公众号二维码
        扫码领资料
        反馈
        TOP
        打开小程序