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Sequencing-related solutions

Betaine, 5M
Ethanol, 70%
Instrument buffer, 1x
Library Storage buffer
PEG 8000, 25%
SBB, 2x
Sinefungin, 10mM
SSC, 20x
STE, 10x
TE, 10x

See also Preparation of solutions on zbio.net.  Comments





    Betaine, 5M, ρ~1.073g/ml

    Betaine, store solid at +4°C, solution — at -20°C

    monohydrate C5H11NO2xH2O; Mw=135.2g/M;
    conc.stock100ml200ml700ml
    Betaine5M135.2g/M67.6g135.2g473.2g
    H2O mQ39.74ml79.4ml277.9ml

    anhydrous C5H11NO2; Mw=117.15g/M;
    conc.stock100ml200ml700ml
    Betaine5M 117.15g/M58.58g117.15g410.03g
    H2O mQ48.73ml97.49ml341.08ml

    • dissolve;
    • incubate at 37°C for 1h;
    • filter through 0,2µm filter;
    • prepare 12ml aliquots in 50ml tubes, store at -20°C (liquid at this temperature);



    Ethanol, 70%

    70% Ethanol; store at NT
    conc.stock50ml250ml1L
    Ethanol70% (v/v)100%, 0.79g/ml27.7g138g553g
    H2O mQ15ml75ml300ml


    Instrument buffer, 1x

    1x Instrument buffer, store at 4°C for 10 days
    density1L
    H2O1g/ml700ml
    10x Instrument buffer1.014g/ml100ml / 101.4g /
    Glycerol1.262g/ml200ml / 252.4g /


    Library Storage buffer

    store at -20°C;
    conc.stock1ml5ml
    Tween0.1%10%10µl50µl
    EB  0.99ml0.95ml


    PEG 8000, 25%

    • prepare ~100mg aliquots & store them at NT;
    • do not store solution;
    conc.stock400µl
    PEG-800025% (w/v)solid100mg
    H2O mQ314µl


    SBB, 2x

    2x Streptavidin Binding Buffer, store at +4°C in ~1ml aliquots
    conc.stock0.5ml1ml12ml
    Tris-HCl, pH 7.510mM1M5µl10µl120µl
    NaCl2M5M200µl400µl4.8ml
    EDTA, pH 81mM0.5M1µl2µl24µl
    H2O mQ294µl588µl7.06ml


    Sinefungin, 10mM

    • prepare ~1mg aliquots & store them at +4°C;
    • store solution at -20°C;
    conc.stock  
    Sinefungin10mM381.4g/M1mg
    H2O mQ260µl


    SSC, 20x, ρ = 1.103

    pH 7.0, store at NT
    conc.stock0.5L1.5L2L
    NaCl3M58.44g/M87.66g263.0g350.6g
    Na3C6H5O7x 5.5H2O0.3M357.1g/M53.57g160.7g214.3g
    H2O mQ440.3ml1.32L1.76L



    STE, 10x

    pH7.5, store at 4°C
    conc.stock1ml5ml
    NaCl1M5M0.2ml1.0ml
    Tris Cl, pH7.50.1M1M0.1ml0.5ml
    EDTA10mM0.5M20µl0.1ml
    H2O mQ680µl3.4ml
    sterilize by autoclaving


    TE, 10x

    pH7.5, store at 4°C
    conc.stock1ml5ml
    Tris Cl, pH7.50.1M1M0.1ml0.5ml
    EDTA10mM0.5M20µl0.1ml
    H2O mQ880µl4.4ml
    sterilize by autoclaving





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