This is a cached page for the URL (http://molbiol.ru/eng/protocol/01_07.html). To see the most recent version of this page, please click here.
Protocol Online is not affiliated with the authors of this page nor responsible for its content.
About Cache
   Compound solutions | molbiol.ru   
molbiol.ru - main page.
 > Protocols >  Solutions > Compound solutions.

Compound solutions.

Rambler's Top100








    About the recalculation of recipes for the arbitrary volume:

    TE

    1x; pH7.4; 7.6; 8.0; (store at 4oC).

    Conc.Stock1ml5ml10ml50ml400ml
    Tris Cl10mM1M10µl50µl100µl500µl4ml
    EDTA1mM0.5M2µl10µl20µl100µl800µl
    H2O mQ988µl4.94ml9.88ml49.4ml395ml

    10x:

    Conc.Stock1ml5ml
    Tris Cl0.1M1M0.1ml0.5ml
    EDTA10mM0.5M20µl0.1ml
    H2OmQ 880µl4.4ml
  • autoclave sterilization
  • STE

    1x; (store at 4oC).

    Conc.Stock1ml5ml10ml50ml
    NaCl0.1M5M20µl100µl200µl1.0ml
    Tris Cl, pH8.010mM1M10µl50µl100µl0.5ml
    EDTA, pH8.01mM0.5M2µl10µl20µl100µl
    H2O mQ0.968ml4.84ml9.68ml48.4ml

    10x:

    Conc.Stock1ml5ml
    NaCl1.0M5M0.2ml1.0ml
    Tris Cl0.1M1M0.1ml0.5ml
    EDTA10mM0.5mM20µl0.1ml
    H2OmQ 680µl3.4ml
  • autoclave sterilization
  • SSC

    20x; 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

    p = 1.103

    TAE

    1x pH = 7.6:

    Tris-acetate 40mM
    EDTA 1mM

    store the large stock at 4oC, working quantity (~100ml) at NT

    Conc.Stock100ml150ml200ml250ml
    Tris-base2M121.1g/M24.22g36.33g48.44g60.55g
    EDTA0.05M372.3g/M1.862g2.792g3.723g4.654g
    Acetic acid~1.56M17.4M ~8.96ml
    ~9.40g
    ~13.44ml
    ~14.10g
    ~17.92ml
    ~18.80g
    ~22.40ml
    ~23.50g
    ~
    ~
    H2O mQ~73.30g~109.95g~146.60g~183.25g~

    p(50x)=1.0878

    it is convenient to solubilize Tris and EDTA first, and only then add acid to adjust pH.

    TBE

    10x, pH ~8.3;(store at NT in clean bottle)

    1x10xStock1L
    Tris base89mM0.89M121.14g/M107.8g
    Boric acid89mM0.89M61.83g/M55.03g
    EDTA, pH 8.02mM20mM500mM40ml
    43.84g

    H2O  mQ863.3ml
  • 10x stock solution is stable at NT if to clean it through 0.4µm filter;
  • for PAAG it is used as 1x, for agarose gel - as 0.5x;
  • for agarose gels it is used for separation of small (< 200bp) fragments. Generally accepted, that the purification of fragments is more complex from TBE gels.
  • FGRB

    50x (formaldehyde gel-running buffer), p=1.104g/ml (store at +4oC):

    Conc.Stock0.5L1.0L2.0L
    MOPS (pH7.0)1.0M209.27g/M104.64g209.27g418.54g
    AcONax3H2O0.5M136.08g/M34.02g68.04g136.08g
    EDTAx2H2O50mM372.3g/M9.31g18.62g37.23g
    H2O mQ404.0ml808ml1616ml
    Conc.Stock0.5L1.0L2.0L
    MOPS (pH7.0)1.0M209.27g/M104.64g209.27g418.54g
    AcONa anhydrous0.5M82.04g/M20.51g41.02g82.04g
    EDTA x 2H2O50mM372.3g/M9.31g18.62g37.23g
    H2O mQ417.5ml835ml1670ml
  • adjust pH7.0 10N NaOH (per 500ml of buffer ~22ml), filter-sterilize through 0.22µm filter;
  • solution becomes yellow during the storage on the light, but it has no effect on the solution quality.
  • GTGB

    20x /Glycerol tolerant gel buffer/ (store at +4oC):

    Conc.Stock1L1.5L2.0L2.5L

    Tris base

    1.78M121.14g/M216g324g432g540g

    Taurine

    0.56M125.15g/M72g108g144g180g

    Na2EDTAx2H2O

    10.7mM372.3 g/M4g6g8g10g

    H2O

     mQ802ml1203ml1604ml2005ml

    p=1.094

  • Taurine: C2H7NO3S; Mw=125.15g/M.
  • TGB

    Tris-Glycine buffer 5x, pH8.3, (store the main stock at +4oC, working quantity at NT).

    1xConc.Stock1L1.5L
    Tris-base25mM125mM121.1g/M15.1g22.71g
    Glycine250mM1.25M75.05g/M94g141g
    SDS0.1%0.5%10%50ml75ml
    H2O  mQ   

    Loading buffer

    10x, (store at NT).

    Conc.Stock1ml5ml10ml25ml
    Xilene cyanol0.1%solid1.0mg5.0mg10mg25.0mg
    Bromph. blue0.1%solid1.0mg5.0mg10mg25.0mg
    SDS0.5%10%50µl250µl500µl1.25ml
    EDTA, pH8.00.1M0.5M0.2ml1.0ml2.0ml5.0ml
    Glycerol50%100% 0.5ml
    0.625g
    2.5ml
    3.125g
    5.0ml
    6.25g
    12.5ml
    15.63g

    H2O mQ0.25ml1.25ml2.5ml6.25ml

    PEG-NaCl

    (store at 4oC).

    Conc.Stock50ml100ml150ml200ml
    NaCl1.6M58.44g/M4.675g9.35g14.03g18.70g
    PEG-600013%solid6.50g13.00g19.50g26.00g

    or:

    Conc.Stock50ml100ml150ml200ml
    NaCl1.6M5M 16ml
    18.97g
    32ml
    37.95g
    48ml
    56.92g
    64ml
    75.90g

    PEG-600013%40%16.25ml32.50ml48.75ml65.00ml

    SM

    (store at 4oC).

    Conc.Stock10ml50ml100ml150ml
    NaCl0.1M5M200µl1.0ml2.0ml3.0ml
    MgSO4*10mM1M100µl0.5ml1.0ml1.5ml
    Tris Cl, pH7.550mM1M0.5ml2.5ml5.0ml7.5ml
    0.01%2%50µl0.25ml0.5ml0.75ml
    H2O mQ9.15ml45.75ml91.5ml137.25ml

    * - add after autoclaving;

  • autoclave sterilization
  • STM

    (store at 4oC):

    Conc.Stock50ml
    NaCl10mM5M100µl
    Tris Cl pH8.050mM1M2.5ml
    MgCl210mM1M500µl
    H2O mQ46.9ml

    TM

    (store at 4oC):

    Conc.Stock50ml
    Tris Cl pH8.050mM1M2.5ml
    MgCl210mM1M500µl
    H2O mQ47.0ml

    Denaturation buffer

    (store at NT).

    Conc.Stock100ml150ml300ml500ml750ml
    NaCl1.5M58.44g/M8.766g13.15g26.30g43.83g65.75g
    NaOH0.5M10M 5ml
    6.65g
    7.5ml
    9.97g
    15ml
    19.94g
    25ml
    33.23g
    37.5ml
    49.84g

      40g/M2g3g6g10g15g

    Neutralization buffer

    (store at NT).

    Conc.Stock150ml300ml500ml750ml
    Tris Cl, pH7.41M121.1g/M18.15g36.3g60.5g90.75g
    NaCl1.5M58.44g/M13.15g26.30g43.83g65.75g
    HCl~0.81M11.6M/L~10.5ml~21ml~35ml~52.5ml
    EDTA (optional)1mM0.5M0.3ml0.6ml1ml1.5ml

    Developer Rentgen-2

    (store at NT in the dark).

    500ml800ml1l
    1.1g1.76g2.2g
    Na2SO3 anhidr.36.0g57.6g72.0g
    Hydroquinone4.4g7.04g8.8g
    Na2CO324.0g38.4g48.0g
    KBr2.0g3.2g4.0g
    H2O490ml784ml980ml

    solubilize in the presented order , p=1.115g/ml.

    Fixer BKF-2

    (store at NT).

    500ml800ml1l
    Na2S2O3x 5H2O130g208g260g
    NH4Cl25g40g50g
    Sodium pyrosulfite8.5g13.6g17g
    H2O398ml636.8ml796ml

    solubilize in the presented order, p=1.123g/ml.

    Scintillation solution

    (store at NT in the dark):

    0.5L1L
    PPO2.50g5.00g
    POPOP0.05g0.10g
    Toluene   

    Plasmid solution I

    (store at +4oC)

    Conc.Stock50ml100ml150ml200ml
    Glucose50mM2M 1.25ml
    1.41g
    2.5ml
    2.825g
    3.75ml
    4.24g
    5.0ml
    5.56g

    Tris Cl, pH 8.025mM1.0 M1.25ml2.5ml3.75ml5.0ml
    EDTA10mM0.5 M1.0ml2.0ml3.0ml4.0ml
    H2O mQ46.5ml93ml139.5ml186ml

    Plasmid solution II

    Conc.Stock0.5ml1ml2.5ml5.0ml6.5ml8.5ml
    NaOH0.2N1N0.1ml0.2ml0.5ml1.0ml1.3ml1.7ml
    SDS1%10%0.05ml0.1ml0.25ml0.5ml0.65ml0.85ml
    H2O mQ0.350ml0.7ml1.75ml3.5ml4.55ml5.95ml

    Oligonucleotide gel staining mix

    (store at NT).

    Conc.Stock100ml200ml300ml500ml
    AcONa(5.2)0.5M3M17ml33ml50.0ml83ml