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Media, vitamins and so on

About the recalculation of recipes for the arbitrary volume:


    Media

    LB, LTB

    pH 7.0; p=1.024



    Stock100ml150ml200ml300ml600ml750ml
    1.Bactotryptonesolid1g1.5g2.0g3.0g6.0g7.5g
    2.Bacto Yeast extr.solid0.5g0.75g1.0g1.5g3.0g3.75g
    3.NaClsolid1.0g1.5g2.0g3.0g6.0g7.5g
    4.H2OmQ95.3g143g190.7g286g572g715g
    5.agar topsolid0.65g0.98g1.3g1.95g3.90g4.88g
    6.agar bot.solid1.5g2.3g3.0g4.5g9.0g11.3g
    7.NaOH10N16.7µl25µl33.4µl50µl100µl125µl
    8.MgSO41M1ml1.5ml2.0ml3.0ml6.0ml7.5ml

    LTB: p.1, 3, 4, 7, 8.
    LB: p.1, 2, 3, 4, 7.
    LB/Mg: p.1, 2, 3, 4, 7, 8.

  • it is recommended to mix agar after autoclaving. Otherwise it will solidify as heterogeneous substance (much denser on the bottom).

  • LB (low salt)

    for using with Zeocine pH 7.0

    Stock100ml150ml200ml300ml600ml750ml
    Bactotryptonesolid1g1.5g2.0g3.0g6.0g7.5g
    Bacto Yeast extr.solid0.5g0.75g1.0g1.5g3.,0g3.75g
    NaClsolid0.5g0.75g1.0g1.5g3.,0g3.75g
    H2OmQ95.3g143g190.7g286g572g715g
    NaOH10N16.7µl25µl33.4µl50µl100µl125µl

    M9

    10x

    Conc.Stock50ml75ml100ml
    Na2HPO4x 12 H2O48mM358.14g/M8.60g12.90g17.19g
    KH2PO422mM136.09g/M1.50g2.25g3.00g
    NaCl8.5mM58.44g/M0.25g0.38g0.50g
    NH4Cl18.7mM53/49g/M0.50g0.75g1.00g

    M9:

    Conc.Stock100ml150ml200ml
    M91x10x10ml15ml20ml
    Glucose0.4%10%4ml6ml8ml
    H2O mQ86ml129ml172ml

    add additional components:

    thiamine1mM1M100µl150µl200µl
    MgSO41mM1M100µl150µl200µl
    CaCl20.1mM1M10µl15µl20µl
  • for making Petri plates add agar to 3%.
  • NZY

    1 L200ml
    NZ amin10g2g
    NaCl5g1g
    Bacto Yeast extr.5g1g
    MgSO4xH2O2g0.4g
    H2O950ml190ml

    SOC, SOB

    (store at NT).

    Stock200ml400ml
    Bactotryptone 4.0g8.0g
    Bacto Yeast extr. 1.1g2.2g
    NaCl5M0.4ml0.8ml
    KCl1M2.0ml4ml
    H2O ~196ml~391ml
    1. add
    2. 0.01V 2M Mg2+{= 1M MgCl2 + 1M MgSO4}  
      0.01V 2M glucose;  

    3. 0.2µm filter sterililize;
    4. store at -20oC, (-70oC) in small aliquots.

    SOB - the same solution without glucose.

    Terrific Broth

    pH 7.0

    100ml150ml200ml300ml667ml
    Bactotryptone1.2g1.8g2.4g3.6g8.0g
    Bacto Yeast extr.2.4g3.6g4.8g7.2g16.0g
    Glycerol0.4ml
    0.504g
    0.6ml
    0.757g
    0.8ml
    1.009g
    1.2ml
    1.513g
    2.667ml
    3.362g

    H2O to V90ml135ml180ml270ml600ml
    after autoclaving, add
    KPO4, pH7.0 1.0M10ml15ml20ml30ml66.7ml

    YENB

    pH 7.5; (non-salt medium for electro-competent cells preparation).

    Conc.Stock0.45 L0.5 L0.9 L1.0 L
    Bacto Yeast extr.0.75%solid3.4g3.75g6.75g7.5g
    Nutrient Broth0.8%solid3.6g4.0g7.2g8.0g
    NaOH 10N~243 µl~270 µl~490 µl~540 µl

    2 YT

    %(w/w)200ml1L
    Bactotryptone1.63.2g16g
    Bacto Yeast extr.1.02g10g
    NaCl0.51g5g
    H2O96.9193.8ml969ml




See also:
/web-links/
  • Difco Manual On-Line (11 issue) - not only recipies, but also papers about the principles of media preparation. It is possible to download the whole manual. /from Renat Gayazov/

    Other substances

    IPTG

    C6H18O5S; Mw=238.3g/M;

    Conc.1ml5ml10ml
    IPTG200mg/ml0.2g1.0g2.0g
  • filter sterililize;
  • store dry powder at 4oC,solution in H2O at -20oC;
  • - for plasmids - add ~4µl per 80mm Petri dish,
    - for M13 - add ~1-4µl per 3ml of top agar.

    X-Gal

    C14H15BrClNO6 Mw= 408g/M;

    Conc.1ml5ml10ml
    X-Gal20mg/ml (2%)20mg0.1g0.2g
  • store dry powder at 4oC in the dark; solution at -20oC in the dark (in DMFA (or DMSO with the same result)).
  • - for plasmids - per 80mm Petri dish 40µl,
    - for M13 - per 3ml of top agar 40µl.

    Glucose

    C6H12O6 Mw=180.16;

    Conc.Stock5ml10ml50ml100ml150ml200ml
    C6H12O61M180g/M0.90g1.80g9.01g18.02g27.03g36.03g
  • filter sterililize.
  • Maltose.

    100x solution for bacteriophage lambda media;

    Conc.Stock10ml50ml100ml
    Maltose20%solid2g10g20g
  • filter sterililize.
  • Thiamine

    Vitamin B1 Aneurine; C12H17ClN4OSxHCl; Mw=337.3g/M;

    Conc.Stock1ml5ml10ml25ml
    C12H17ClN4OSxHCl1M337.3g/M0.337g1.69g3.37g8.43g
  • filter sterililize.


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Last modification: 30/07/13

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