[mouse work] proteinase_K_만들기

This topic has 0 replies, 1 voice, and was last updated 16 years, 1 month ago by Seung Jae Jeong.

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    • #18922

      Seung Jae Jeong
      Participant

      I. proteinase K만들기.

       

       

      stock

      working

      100ml

      Tris-HCl(pH 7.5)

      1M

      10mM

      1ml

      CaCl2

      1M

      20mM

      2ml

      Glycerol

       

      50%

      50ml

      0.22 filtered D.W

       

       

      47ml

      Glycerol treatment 주위 (정확한 넣어줄 .)

       

      100ml solution total 1g PK 녹여준다.

      10mg/ml stock 만들어짐.

       

      II. TES 만들기

        1cm정도로 자른 꼬리에 TES 500+ PK20(=100~200/1cm tail)

            처리한 후 58OC에서 O/N

      .Water bath 58OC로 미리 setting 해놓는다.

      PK60OC 이상으로 처리할 경우,PK가 파괴되므로 절대 넘지 않도록 주의.

      PK는 녹여서 잘 inverting해준 후에 사용한다.

       

       

       

       

       

       

       

       

       

       

       

       

       

       

      II. TES 만들기.

       

       

      Stock

      Working

      40ml

      Tris-HCl (pH8.0)

      1M

      0.05M

      2ml

      EDTA (pH8.0)

      0.5M

      0.05M

      4ml

      SDS

      10%

      0.5%

      2ml

      D.W

       

       

      32ml

       

       

       

       

       

       

       

       

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