DNA extraction of animal tissue

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    • #7782
      arm_chair27
      Participant

      i ever used Intelligene’s lysis buffer. Its’ protocol is very easy. you can put your sample tissue in lysis buffer –>grind it well–>incubate at 95 degree celcius for 15 min–>centrifuge–>pipette the supernatant to 95% ethanol–>centrifuge–>dry pellet and dissolve with TE buffer or DI water.
      After i check DNA quality with spectrophotometry and gel electrophoresis, i’s ok–>there are protein interfere a little. I would like to know what is the component of this lysis buffer. Who can guess? please tell me.thank you. 😀

    • #73623
      mith
      Participant

      Some sort of detergent I’m guessing? In my lab we also add proteinase to it before incubating at 56C for 1-3 hrs.

    • #73630
      canalon
      Participant

      I suspect more than one component. You can find some indication in the MSDS or safety sheets (in case you drink it you need to know what to do before you decide to extract your total DNA.

      But you will probably find a strong detergent, a protein denaturant (GITC for example) and maybe something that will precipitate sugar such as CTAB.

    • #74056
      molecular newbie
      Participant

      usually we add proteinease K which denatures all the proteins….if you try adding pro K along with the lyssis buffer chances are you wont get protein interference…… 8)

    • #74133
      MichaelXY
      Participant

      Geez, I had no idea what you guys were talking about, maybe after next semesters Micro Bio class, but that Pro K sounds like a good agent for jellyfish stings. Sounds better than the alternative, which I am sure you have heard of.

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