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silica spin

, silica particles, , . , DNA.
  1. DNA (100mg=100µl);
  2. 4 AB( NIB);
  3. ~10' 50 ;
  4. pH, ;
  5. , , ;
  6. ( 1 – 2');
  7. 400µl AB( NIB) (optional, );
  8. 400µl WB;
  9. 2ml - 2';

  10. DNA:
  11. 1.5ml ;
  12. ~30µl EB, >1';
  13. 2'.


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    AB (adsorbtion buffer)

    ( 4o):
    Guanidine isothiocianate = GuSCN

    .10.0ml50.0ml
    GuSCN5M118.16g/M5.91g29.54g
    MES, pH 5.550mM1M0.5ml2.5ml
    EDTA, pH8.020mM0.5M0.4ml2.0ml
    Triton X-1000.5%10%0.5ml2.5ml
    Cresol red0.05mM50mM10µl50µl
    H2O mQ4.09ml20.45ml

    MES, pH 5.5, 1M

    ( 4o):

    .10.0ml50.0ml
    MES (Na)1M217.2g/M2.17g10.86g
    . - (.)853mM17.4M0.49ml2.45ml
    H2O mQ   

    WB

    ( NT):

    .10.0ml50.0ml
    80%100%8.0ml40.0ml
    TrisCl, pH7.510mM1M0.1ml0.5ml
    H2O mQ1.9ml9.5ml

    EB

    ( 4o):

    .10.0ml50.0ml
    TrisCl, pH8.510mM1M0.1ml0.5ml
    H2O mQ9.9ml49.5ml


  • silica spin " pDNA silica spin ". , ( ).
  • : GuSCN ( NaI), , GuSCN 80% , DNA ( ).
  • silica spin "silica particles (glass milk)" "diatoms" ( — ).
  • , .

  • . 2.

  • Guanidine isothiocianate, NaI, . NIB: 6.6M NaI, 16mM Na2SO3, 0.05mM Cresol Red ( 4oC)
  • . 4.

  • AB , pH: pH<7.5 pH>7.5. , pH>7. (.1) , pH 3M , pH 4. , .
  • . 8.

  • WB. - 80% . , . Tris — WB, 80% .
  • . 11.

  • EB. pH 7.0 - 8.5. , , , pH .


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: 30/07/13

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