• 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

        Bioanalyzer

        互联网

        1498

         

        Protocol for Bioanalyzer RNA nano chip

        preparation of material

        • 12 samples per chip
        • quantitative range 25�500 ng/μl
        • quantitation accuracy 20%CV (for ladder as sample)

        cleaning, gel preparation (start 40min before experiement)

        • take out filtered gel aliquot and fluorescent dye from fridge next door 30min ahead of time (1 gel aliquot tube enough for 2 chips)
        • take out ladder from fridge upstairs
        • vortex dye 10s and spin down
        • mix one tube gel aliquot with 1µl of dye
        • vortex, centrifuge 15000g +-20% for 10min
        • wash electrodes 1min RNase ZAP, 3min water (pipette into 1 well, will spread)

        prepare chip for measurement

        • take out new RNA chip (pico or nano) from drawer below microarray machine and put into station
        • fill 9µl gel into dark circle G well (gel reservoir)
        • press down plunger and wait 30sec (gel moves through channels)
        • after 30sec release silver trigger (should jump up well above 0.3, meaning seal was tight)
        • wait 5sec, pull plunger up all the way
        • open priming station, add 9µl gel to light circle G wells
        • 5µl of marker into all sample wells and ladder well, bottom right (contains small marker to align plots)
        • 1µl of sample per well, add 1µl of water or replicates into free wells (6µl required per well for machine to run properly)
        • 1µl of ladder into ladder well
        • vortex in holder for 1min and put into machine

        start the run

        • start 2100 expert software
        • choose programme for kit: DNA/RNA pico/RNA nano and material: total RNA/mRNA, prokaryotic/eukaryotic
        • start run

        wash, export data

        • typical RNA nano run takes just over 20min
        • wash with clean water for 3min immediately after the run has ended (electrodes in the lid will quickly deteriorate otherwise)
        • to export data as PDF you have to print ; (PDF option checked will save file, PDF unchecked will print on paper)

        Troubleshooting

        A few typical problems occur once in a while:

        • small RNA marker is not properly (often the next peak is selected => fragments size mislabelled)
        cleak peak table, a tab at the bottom of the window; lower marker can be manually set here
        • 18S, 28S not properly assigned (=> RIN number incorrect)

         

        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