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Division of Molecular Biology

Institute of Medical Biochemistry

Division of Molecular Biology

Research Groups

   

   

 

    Buffers and Solutions

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    100ml 40% Acrylamid in H2O:
    38 g Acrylamid
    2 g NN´Methylenbisacrylamid
    1 spoon DOWEX


    Aminoacid-Mix (for Yeast):
    Amino acids from Sigma

    L-Adenine 0,4 g
    L-Arginine 0,2g
    L-Tyrosine 0,3g
    L-Isoleucine 0,3g
    L-Phenylalanine 0,5g
    L-Glutamic acid 1g
    L-Aspartic acid 1 g
    L-Threonine 2g
    L-Serine 4g
    L-Valine 1,5g
    L- Methionine 1,5g
    L-Lysine 1,8g


    Antibiotics:
    1000 x Ampicillin (50 mg / ml)
    1000 x Chloramphenicol (30 mg / ml)
    1000 x Kanamycin (50 mg / ml)
    1000 x Tetracyclin (30 mg / ml)


    Binding-buffer 2x:
    40 mM HEPES pH=7,9
    2 mM MgCl2
    8 % (w/v) Ficoll or 10-15 % Glycerol
    80 mM KCl
    0,2 mM EDTA
    0,2 % NP-40


    Coomassieblue:
    0,25 % (w/v) Coomassie R
    10 % (v/v) Acetic Acid
    50 % (v/v) Methanol

    DEMEM (10 Liter):
    DEMEM-Pulver in 5 Liter Wasser lösen
    Zugabe von 30 g NaHCO3 (5 min lösen)
    Auffüllen auf 10 Liter.
    Sterilfiltrieren (Satotrius P plus - Filter).
    Je 460 ml aliquotieren.


    Drop Out 2x (for Yeast)[740 ml]:
    3,4 g Bacto-YNB w/o amino acids
    and (NH4)2SO4 (Difco)
    10 g (NH4)2SO4 (Merck)
    2,86 g Amino-Acid-Mix
    X ml H2O
    pH= 5,5 - 6,0


    Trypsin-EDTA (250 ml):
    250 mg Trypsin
    245 ml PBS 10x
    5ml 1% NaEDTA pH=7,5
    steril filtration


    DTT 100mM:
    0,07715g Dithiothreitol / 5ml H2O


    FAX-staining solution:
    100mM Tris-HCl pH= 7,5
    2mM MgCl2
    0,1 % Triton X-100
    2µg / ml DAPI (4,6-Diamidino-2-phenylindolhydrochlorid; Merck Cat.No. 25653)
    15 µg/ml Sulforhodamin 101 (Sigma Cat.No. S-7635)


    Formamid-DNA-Stoppsolution
    (for Sequencing):
    95% Formamid
    20mM EDTA
    0.05% Bromphenol Blue
    0.05% Xylene-Cyanol FF


    FRAKELTON-Buffer:
    10 mM Tris-HCl
    50 mM NaCl
    1 % Triton X-100
    30 mM Na-Pi
    50 mM NaF
    2 µM ZnCl2
    10 mM Na3VO4
    (PMSF, DTT.)
    (Na3VO4 in H2O pH=9,5).


    2x Freezing buffer (1000ml):
    12,6 g K2HPO4
    3,6 g KH2PO4
    0,9 g Na-Citrat
    0,18 g MgSO4 x 7 H2O
    1,8 g (NH4)2SO4
    88 ml Glycerol


    Glycogen (for DNA Precipitation):
    20 mg/ml


    HBS 2x:
    280 mM NaCl
    50 mM HEPES
    1,5 mM Na2HPO4 x 12 H2O
    pH=7,4
    steril filtration


    Histidin 100x (for Yeast):
    0,6 g L-Histidin in 100 ml H2O


    HUNT-Buffer:
    20 mM Tris / HCl pH=8,0
    100 mM NaCl
    1 mM EDTA
    0,5 % NP-40


    Kinase-Assay buffer:
    50 mM HEPES pH=7,5
    10 mM MgCl2
    1 mM DTT
    [25 µM ATP]


    IPTG 100mM:
    23,8 mg Isoprpyl-ß-D-thio-galactopyranoside / ml H2O


    dNTP-Mix 10x:
    2 mM dATP
    2 mM dCTP
    2 mM dGTP
    2 mM dTTP


    Leucin 100x (for Yeast):
    2,6 g L-Leucin in 100 ml H2O


    Luciferase-assay:

    Luciferaseassaybuffer:
    25 mM Glycylglycin pH 7,8
    15 mM Kaliumphosphat pH 7,8
    15 mM MgSO4
    4 mM EGTA
    3,4 mM ATP (Sigma # L3377)

    Triton-Glycylglycin buffer:
    1 % (v/v) Triton X-100
    25 mM Glycylglycin pH 7,8
    15 mM MgSO4
    4 mM EGTA
    1 mM DTT

    Luciferin:
    800 µM D-Luziferin (Sigma # L9504)
    25 mM Glycylglycin pH 7,8
    10 mM DTT

    Luciferase:
    1 ng (1 : 10E4 of 1 mg / ml stock)


    10xPBS [250ml] pH=7,3:
    1,4 M NaCl (20,454g)
    0,027 M KCL (0,5033g)
    0,1 M Na2HPO4 (3,55g)
    0,018 M K2HPO4 (3,55g)
    0,018 M KH2PO4 (0,612g)


    Phenol:
    Add to Phenol 0,1% W/V Hydroxyquinoline
    Extract three times with 0,1 M Tris pH=7,8 - 8,0
    Add an equal volume of 0,01 M Tris pH= 7,8 - 8,0


    Phenol & Chloroform:
    25 parts Phenol + 0,1 % W/V Hydroxyquinoline
    24 parts Chloroform
    1 part Isoamyl alcohol
    Mix, then extract several times with 0,1 M Tris pH=7,6 - 8,0
    Add an equal volume of 0,01 M Tris pH=7,6 - 8,0


    Phosphatebuffer:

    57,7 ml 1M Na2HPO4 (35,5g / 250 ml)
    42,3 ml 1M NaH2PO4 (29,995g / 250 ml)


    PMSF 100 mM:
    Dissolve 0,1743 g Phenylmethylsulfonylfluoride in 10 ml 96 % Ethanol.


    Ponceau:
    0,2 % w/v Ponceau S in 3 % Trichlor-acetic-acid


    Protein sample buffer:
    100 mM Tris-HCl pH=6,8
    20 % Glyzerin
    0,01 % Bromphenolblue
    10 % ß-Mercaptoethanol
    5 % SDS
    Dissolve in H2O


    Running buffer 1x:
    192 mM Glycin
    25 mM Tris
    0,1 % SDS
    Dissolve in H2O


    Stripping-buffer:
    100 mM 2-Mercaptoethanol
    2 % (w/v) SDS
    62,5 mM Tris-HCl
    pH 6,7


    Southernblot-Solutions:

    Neutralisation Solution (1000ml):
    200 ml 20 x SSC
    100 ml 1N Hcl
    100ml 1M Tris-HCl pH=8
    X ml H2O

    20 x SSC:
    3M NaCl
    0,3 M Na3Citrat

    Wallace:
    30 ml 10 x NET
    5 ml 50 x Denhardts
    2,5 ml 10 % SDS
    250 µl 10 mg / ml tRNA

    10 x NET:
    1,5 M NaCl
    150 mM Tris-HCl pH=7,5
    10 mM EDTA

    50 x Denhardts:
    1 % BSA
    1 % Ficoll
    1 % PVP (Polyvinylpyrrolidon)


    TAE 50x (1000ml):
    242g Tris Base
    57,1g Glacial Acetic Acid
    100ml 0,5M EDTA


    TBE 20x (1000ml):
    216g Tris Base
    110g Borat
    80ml 0,5M EDTA pH=8


    TBST 1x:
    10 mM Tris pH=8
    150 mM NaCl
    0,1 % Tween


    TE:
    10 mM Tris-HCl pH=8
    1 mM EDTA pH=8


    TNE-Extractionbuffer:
    10 mM Tris-HCl pH=8
    10 mM NaCl
    0,5 mM EDTA

    1% NP-40


    Tryptophan 100x (for Yeast):
    0,8 g L-Tryptophane in 100 ml H2O
    (steril filter)


    Ultra New Wash:
    50 mM NaCl
    10 mM Tris/HCl pH=7,5
    25mM EDTA
    50 % v/v Ethanol


    Uracil 100x (for Yeast):
    0,4 g L-Uracil in 100 ml H2O


    WCE-Buffer:
    20 mM Hepes pH 7,9
    400 mM NaCl
    25 % v/v Glycerol
    1mM EDTA


    1x Western-Transfer-Buffer (8 Liter):
    1,6 l Methanol
    115,6 g Glycin
    24,2 g Tris
    X liter H2O

    X-Gal:
    40mg / ml in Dimethylformamid


    Z-Buffer:
    Na2HPO4 x 7H2O 16,1 g
    NaH2PO4 x 1H2O 5,5 g
    KCl 0,75 g
    MgSO4 x 7H2O 0,25 g
    2-Mercaptoethanol 2,7 ml
    pH=7


  • Acrylamid:
  • Aminoacid-Mix:
  • Antibiotics:
  • Binding-buffer 2x:
  • Coomassieblue:
  • DEMEM (10 Liter):
  • Drop Out 2x (for Yeast)[740 ml]:
  • Trypsin-EDTA (250 ml):
  • DTT 100mM:
  • FAX-staining solution:
  • Formamid-DNA-Stoppsolution
    (for Sequencing):
  • FRAKELTON-Buffer:
  • 2x Freezing buffer (1000ml):
  • Glycogen:
  • HBS 2x:
  • Histidin 100x (for Yeast):
  • HUNT-Buffer:
  • Kinase-Assay buffer:
  • IPTG 100mM:
  • dNTP-Mix 10x:
  • Leucin 100x (for Yeast):
  • Luciferase-assay:
  • 10xPBS [250ml] pH=7,3:
  • Phenol:
  • Phenol & Chloroform:
  • Phosphatebuffer:
  • PMSF 100 mM:
  • Ponceau:
  • Protein sample buffer:
  • Running buffer 1x:
  • Stripping-buffer:
  • Southernblot-Solutions:
  • TAE 50x (1000ml):
  • TBE 20x (1000ml):
  • TBST 1x:
  • TE:
  • TNE-Extractionbuffer:
  • Tryptophan 100x (for Yeast):
  • Ultra New Wash:
  • Uracil 100x (for Yeast):
  • WCE-Buffer:
  • 1x Western-Transfer-Buffer (8 Liter):
  • X-Gal:
  • Z-Buffer:
 

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