Science Gateway > Protocols > Cell Biology Protocols - Table of Contents

Pubplus.org - a PubMed search engine
Star Republic: Guide for Biologists

Recipes

ATP (adenosine triphosphate 10 mM)
disodium ATP0.055gram (g)
Sterile ddH2O to10millilitre (ml)
Total volume10millilitre (ml)
Note: aliquot and store at -20 degree

Acrylamide/Bisacrylamide (19:1, 40%)
Acrylamide380gram (g)
N,N-Methylene-bisacrylamide 20gram (g)
ddH2O to1litre (l)
Total volume1litre (l)
Note: filter and store protected from light at 4 degree

Acrylamide/Bisacrylamide (37.5:1, 30%)
acrylamide300gram (g)
bisacrylamide8gram (g)
ddH2O to1litre (l)
Total volume1litre (l)
Note: filter and store protected from light at 4 degree

Agarose gel loading dye (10X)
Ficoll1.5gram (g)
Bromophenol blue 0.02gram (g)
xylene cyanole FF 0.02gram (g)
ddH2O to10millilitre (ml)
Total volume10millilitre (ml)

Alkaline lysis solution (NaOH/SDS)
1N NaOH20millilitre (ml)
10% SDS10millilitre (ml)
ddH2O to100millilitre (ml)
Total volume100millilitre (ml)

Ammonium acetate (5M)
NH4OAc385.4gram (g)
ddH2O to500millilitre (ml)
Total volume500millilitre (ml)
Note: filter to sterilize

Ammonium acetate (8.0M)
NH4OAc61.7gram (g)
ddH2O to100millilitre (ml)
Total volume100millilitre (ml)

Ammonium acetate (9.5M)
NH4OAc72.3gram (g)
ddH2O to100millilitre (ml)
Total volume100millilitre (ml)

Ammonium persulfate (10%)
Ammonium persulfate 1gram (g)
ddH2O to10millilitre (ml)
Total volume10millilitre (ml)

Ampicillin (50 mg/ml)
Ampicillin 0.5gram (g)
Sterile ddH2O to10millilitre (ml)
Total volume10millilitre (ml)
Note: aliquot and store at -20 degree

BME (beta-mercaptoethanl 0.5M)
beta-mercaptoethanl3.5millilitre (ml)
ddH2O96.5millilitre (ml)
Total volume100millilitre (ml)

BME (beta-mercaptoethanl 1M)
beta-mercaptoethanl6.98millilitre (ml)
ddH2O93millilitre (ml)
Total volume100millilitre (ml)

BSA (1mg/ml)
BSA5milligram (mg)
Sterile ddH2O to5millilitre (ml)
Total volume5millilitre (ml)
Note: aliquot and store at -20 degree

Calcium chloride (CaCl2)
CaCl2*2H2O1.48gram (g)
ddH2O to100millilitre (ml)
Total volume100millilitre (ml)
Note: autoclave to sterilize and store at 4degree

Chloramphenicol (25 mg/ml)
Chloramphenicol250milligram (mg)
Ethanol (100%)10millilitre (ml)
Total volume10millilitre (ml)
Note: dissolve in absolute ethanol, aliquot and store at -20 degree

DTT (Dithiothreitol 1M)
DTT1.54gram (g)
ddH2O to10millilitre (ml)
Total volume10millilitre (ml)
Note: aliquot and store at -20 degree

Denhart Solution (100X)
BSA2gram (g)
Ficol 400,0002gram (g)
Polyvinylpyrrolidone 40,0002gram (g)
SSPE (20X)15millilitre (ml)
ddH2O to100millilitre (ml)
Total volume100millilitre (ml)
Note: store at -20 degree

EDTA (0.5M pH8.0)
Na2EDTA186.1gram (g)
ddH2O to500millilitre (ml)
Total volume1000millilitre (ml)
Note: Dissolve in approx. 400 ml ddH2O, adjust pH to 8.0 with 10 N NaOH, and adjust to 1 liter final volume with distilled water. Autoclave to sterilize

EtBr (ethidium bromide 5 mg/ml)
Ethidium bromide500milligram (mg)
ddH2O to100millilitre (ml)
Total volume100millilitre (ml)

Ethonal (70%)
95% Ethonal74millilitre (ml)
ddH2O26millilitre (ml)
Total volume100millilitre (ml)

Gelatin (2%)
gelatin2gram (g)
ddH2O to100millilitre (ml)
Total volume100millilitre (ml)
Note: autoclave and store at room temperature.

Glucose (1M)
glucose18gram (g)
ddH2O to100millilitre (ml)
Total volume100millilitre (ml)
Note: filter to sterilize

Glucose (20%)
Glucose20gram (g)
ddH2O to100millilitre (ml)
Total volume100millilitre (ml)
Note: filter sterilize

Glycerol (Sterile)
Note: autoclave to sterilize.

Guanidine (6M pH6.4)
Guanidine-HCl573.18gram (g)
1M Tris-HCl (pH7.6)50millilitre (ml)
0.5 M EDTA40millilitre (ml)
ddH2O to1litre (l)
Total volume1litre (l)

HEPES (1M pH7.5)
HEPES23.83gram (g)
ddH2O to100millilitre (ml)
Total volume100millilitre (ml)
Note: adjust pH to 7.5 with potassium hydroxide (KOH) and store at 4 degree

Hygromycin B (100 mg/ml)
Hygromycin1gram (g)
ddH2O to10millilitre (ml)
Total volume10millilitre (ml)
Note: filter sterilize, aliquot and store at -20 degree

IPTG (20mg/ml)
IPTG200milligram (mg)
ddH2O10millilitre (ml)
Total volume10millilitre (ml)
Note: filter sterilize, aliquot and store at -20 degree

IPTG (isopropyl b-D-thiogalactopyranoside 25 mg/ml)
IPTG250milligram (mg)
ddH2O to10millilitre (ml)
Total volume10millilitre (ml)

Isocitrate (1M)
Na3isocitrate-2H2O 29.41gram (g)
ddH2O to100millilitre (ml)
Total volume100millilitre (ml)

KCl (1M)
KCl7.5gram (g)
ddH2O to100millilitre (ml)
Total volume100millilitre (ml)
Note: autoclave to sterilize

Kanamycin (25mg/ml)
Kanamycin sulfate 0.25gram (g)
Sterile ddH2O to10millilitre (ml)
Total volume10millilitre (ml)
Note: aliquot and store at -20 degree

LB medium
Bacto-Tryptone 10gram (g)
Bacto-yeast extract 5gram (g)
NaCl10gram (g)
ddH2O to1litre (l)
Total volume1litre (l)
Note: adjust pH to 7.0 and autoclave to sterilize

LB plates
Bacto-Tryptone 10gram (g)
Bacto-yeast extract 5gram (g)
NaCl10gram (g)
Bacto-agar 15gram (g)
ddH2O to1litre (l)
Total volume1litre (l)
Note: autoclave to sterilize, cool to 55 degree, add antibiotic if desired, and pour into sterile petri dishes

Lambda plates
Bacto-tryptone 10gram (g)
Bacto-agar 15gram (g)
NaCl2.5gram (g)
ddH2O to1litre (l)
Total volume1litre (l)
Note: autoclave to sterilize and pour into sterile petri dishes

Lambda top agar
Bacto-tryptone 10gram (g)
Bacto-agar 10gram (g)
NaCl5gram (g)
ddH2O to1litre (l)
Total volume1litre (l)
Note: autoclave to sterilize

M9 medium
Na2HPO46gram (g)
KH2PO43gram (g)
NaCl0.5gram (g)
NH4Cl1gram (g)
ddH2O to1litre (l)
Total volume1litre (l)
Note: autoclave. Add 10 ml filter sterilized 100 mM MgSO4, 20% glucose, 10 mM CaCl2, 100 mM thiamine-HCl) before use.

MOPS (1M)
MOPS20.9gram (g)
ddH2O to100millilitre (ml)
Total volume100millilitre (ml)
Note: Dissolve in 80 ml ddH2O, adjust pH to 7.5 with 1 N NaOH, and bring volume to 100 ml with ddH2O

Magnesium chloride (1M)
MgCl2*6H2O20.3gram (g)
ddH2O to100millilitre (ml)
Total volume100millilitre (ml)
Note: autoclave to sterilize

Magnesium sulfate (1M)
MgSO412gram (g)
ddH2O to100millilitre (ml)
Total volume100millilitre (ml)
Note: autoclave to sterilize

Manganese chloride (1M)
MnCl21.98gram (g)
ddH2O to10millilitre (ml)
Total volume10millilitre (ml)
Note: store protected from light

NYZ agar
NaCl5gram (g)
MgSO4*7H2O2gram (g)
Bacto-yeast extract 5gram (g)
NZ amine (casein hydrolysate)10gram (g)
Bacto-agar15gram (g)
ddH2O to1litre (l)
Total volume1litre (l)

NYZ top agarose
NYZ Broth1litre (l)
agarose7gram (g)
Total volume1litre (l)
Note: autoclave

NZY Broth
NaCl5gram (g)
MgSO4*7H2O2gram (g)
Bacto-yeast extract 5gram (g)
NZ amine (casein hydrolysate)10gram (g)
ddH2O to1litre (l)
Total volume1litre (l)
Note: adjust pH to 7.5 and autoclave

Neutral Buffered Formalin
Formalin (40% formadehyde)100millilitre (ml)
Sodium phosphate, monobasic, monohydrate4gram (g)
Sodium phosphate, dibasic, anhydrous6.5gram (g)
ddH2O to1litre (l)
Total volume1litre (l)

P1 (15 mM Tris pH8 10mM EDTA plasmid prep)
1M Tris-HCl (pH 8.0)1.5millilitre (ml)
0.5M EDTA2millilitre (ml)
ddH2O to96.5millilitre (ml)
Rnase A10milligram (mg)
Total volume100millilitre (ml)
Note: filter sterilize, or autoclave and add Rnase A before use.

P2 (0.2N NaOH 1% SDS plasmid prep)
10N NaOH2millilitre (ml)
10% SDS10millilitre (ml)
ddH2O88millilitre (ml)
Total volume100millilitre (ml)
Note: autoclave to sterilize

P3 (3M KOAc pH 5.5 plasmid prep)
KOAc54gram (g)
ddH2O to100millilitre (ml)
Total volume100millilitre (ml)
Note: dissolve in 80 ml of ddH2O, adjust to pH5.5 with glacial acetic acid and add ddH2O to 100 millilitre. Autoclave to sterilize

Phenol (TE-saturated)
Note: add an equal volume of TE (pH7.5-8.0) to phenol, mix well, allow phases to allow phases to separate, remove and discard upper (aqueous) phase. Repeat until the pH of the aqueous phase is between 7.5-8.0 (store protected from light at 4 degree)
chloroform/isoamylalcohol(25:25:1)>

Phenol/chloroform/isoamyl alcohol (25:25:1)
TE-saturated phenol100millilitre (ml)
chloroform100millilitre (ml)
isoamyl alcohol4millilitre (ml)
Total volume204millilitre (ml)
Note: store protected from light at 4 degree

Polyacrylamide Gel loading dye
xylene cyanole FF3gram (g)
bromophenol blue3gram (g)
0.5 M EDTA0.2millilitre (ml)
deionized formamide 10millilitre (ml)
Total volume100millilitre (ml)
Note: for sequencing gel

Potassium acetate (7.5M)
KOAc736gram (g)
ddH2O to1litre (l)
Total volume1litre (l)

Proteinase K (10mg/ml)
Proteinase K0.01gram (g)
TE1millilitre (ml)
Total volume1millilitre (ml)
Note: aliquot and store at -20 degree

SM
NaCl5.8gram (g)
MgSO4*7H2O2gram (g)
1M Tris-HCl pH7.550millilitre (ml)
Gelatin (2%)5millilitre (ml)
ddH2O to1litre (l)
Total volume1litre (l)
Note: autoclave and store at room temperature.

SOB medium
Bacto-tryptone 20gram (g)
Bacto-yeast extract 5gram (g)
NaCl0.5gram (g)
1M KCl2.5millilitre (ml)
ddH2O to1000millilitre (ml)
Total volume1000millilitre (ml)
Note: adjust pH to 7.0 with 10N NaOH, autoclave to sterilize, and add 10 ml of 1 M MgCl2 before use

SOC medium
Bacto-tryptone 20gram (g)
Bacto-yeast extract 5gram (g)
NaCl0.5gram (g)
1M KCl2.5millilitre (ml)
ddH2O to1000millilitre (ml)
Total volume1000millilitre (ml)
Note: adjust pH to 7.0 with 10N NaOH, autoclave to sterilize, add 20 ml of sterile 1 M glucose immediately before use

SSC (20X)
NaCl175.3gram (g)
Sodium citrate88.22gram (g)
ddH2O to1000millilitre (ml)
Total volume1000millilitre (ml)
Note: dissolve in 800 ml of ddH2O, adjust to pH7 with HCl and add ddH2O to 1 litre

SSPE (20X)
NaCl310.4gram (g)
1M phosphate buffer200millilitre (ml)
0.5 M EDTA40millilitre (ml)
ddH2O to1litre (l)
Total volume1litre (l)
Note: autoclave and store at room temperature.

Sodium Acetate (2M)
NaOAc*3H2O27.22gram (g)
ddH2O to100millilitre (ml)
Total volume100millilitre (ml)

Sodium Acetate (3M pH4.8)
NaOAc*3H2O40.8gram (g)
ddH2O to100millilitre (ml)
Total volume100millilitre (ml)
Note: dissolve in 80 ml of ddH2O, adjust to pH4.8 with glacial acetic acid and add ddH2O to 100 millilitre. Filter to sterilize

Sodium Acetate (3M pH5.2)
NaOAc*3H2O40.8gram (g)
ddH2O to100millilitre (ml)
Total volume100millilitre (ml)
Note: dissolve in 80 ml of ddH2O, adjust to pH5.2 with glacial acetic acid and add ddH2O to 100 millilitre. Filter to sterilize

Sodium chloride (3M)
NaCl17.5gram (g)
ddH2O to100millilitre (ml)
Total volume100millilitre (ml)

Sodium chloride (5M)
NaCl292.2gram (g)
ddH2O to1000millilitre (ml)
Total volume1000millilitre (ml)
Note: autoclave to sterilize

Sodium hydroxide (10N)
NaOH40gram (g)
ddH2O to100millilitre (ml)
Total volume100millilitre (ml)

Sodium hydroxide (1N)
10 N NaOH10millilitre (ml)
ddH2O90millilitre (ml)
Total volume100millilitre (ml)

Sodium phosphate (pH6.7)
monobasic sodium phosphate94.5gram (g)
dibasic sodium phosphate44.7gram (g)
ddH2O to1litre (l)
Total volume1litre (l)
Note: filter to sterilize

Spermidine (0.1M)
Spermidine0.255gram (g)
ddH2O to10millilitre (ml)
Total volume10millilitre (ml)
Note: store at -20 degree

T-TYN agar
T-TYN broth1litre (l)
Bacto-agar15gram (g)
Total volume1litre (l)
Note: autoclave.

T-TYN broth
Bacto-yeast extract 5gram (g)
Bacto-tryptone 10gram (g)
NaCl5gram (g)
1M Tris-HCl pH7.210millilitre (ml)
ddH2O to1litre (l)
Total volume1litre (l)
Note: adjust pH to 7.2. autoclave

TBE (10X)
Tris base216gram (g)
Boric Acid110gram (g)
EDTA16.6gram (g)
Water to2litre (l)
Total volume2litre (l)

TE Buffer (100:1)
1 M Tris-HCl (pH 8.0)1millilitre (ml)
0.5 M EDTA0.02millilitre (ml)
Water to100millilitre (ml)
Total volume100millilitre (ml)

TE Buffer (10:1)
1 M Tris-HCl (pH 8.0)1millilitre (ml)
0.5 M EDTA0.2millilitre (ml)
Water to100millilitre (ml)
Total volume100millilitre (ml)

Tetracycline (10 mg/ml)
Tetracycline1gram (g)
Ethanol (100%)50millilitre (ml)
Sterile ddH2O to100millilitre (ml)
Total volume100millilitre (ml)
Note: dissolve in 50% ethonal, aliquot and store protected from light at -20 degree

Tris-HCl (1M)
Tris base121.1gram (g)
ddH2O to1000millilitre (ml)
Total volume1000millilitre (ml)
Note: dissolve in 800 ml of ddH2O, adjust to desired pH with concentrated HCl and add ddH2O to 1 litre

Tris-HCl (2M)
Tris base242.2gram (g)
ddH2O to1litre (l)
Total volume1litre (l)
Note: dissolve in 800 ml of ddH2O, adjust to desired pH with concentrated HCl and add ddH2O to 1 litre

Tris-acetate-EDTA buffer (50x TAE)
Tris-HCl240gram (g)
Glacial acetic acid57.1millilitre (ml)
0.5 M EDTA10millilitre (ml)
Water to1000millilitre (ml)
Total volume1000millilitre (ml)
Note: Store at room temperature

Tris-glycine Native-Polyacrylamide Gel loading buffer (2X)
1M Tris-HCl (phH6.8)1.6millilitre (ml)
Glycerol (100%)2millilitre (ml)
B-mercaptoethanol1millilitre (ml)
Bromophenol blue4milligram (mg)
ddH2O5.4millilitre (ml)
Total volume10millilitre (ml)
Note: store at 4 degree

Tris-glycine Native-Polyacrylamide Gel running buffer (10X)
Tris base30.3gram (g)
Glycine144gram (g)
ddH2O to1litre (l)
Total volume1litre (l)

Tris-glycine SDS-Polyacrylamide Gel (10% Separating)
30% Polyacrylamide33.3millilitre (ml)
1.5M Tris(pH8.8)25millilitre (ml)
Ammonium persulfate (10%)1millilitre (ml)
SDS (10%)1millilitre (ml)
TEMED0.04millilitre (ml)
dH2O39.7millilitre (ml)
Total volume100millilitre (ml)
Note: use immediately

Tris-glycine SDS-Polyacrylamide Gel (12% Separating)
30% Polyacrylamide40millilitre (ml)
1.5M Tris(pH8.8)25millilitre (ml)
Ammonium persulfate (10%)1millilitre (ml)
SDS (10%)1millilitre (ml)
TEMED0.04millilitre (ml)
dH2O33millilitre (ml)
Total volume100millilitre (ml)
Note: use immediately

Tris-glycine SDS-Polyacrylamide Gel (15% Separating)
30% Polyacrylamide50millilitre (ml)
1.5M Tris(pH8.8)25millilitre (ml)
Ammonium persulfate (10%)1millilitre (ml)
SDS (10%)1millilitre (ml)
TEMED0.04millilitre (ml)
dH2O23millilitre (ml)
Total volume100millilitre (ml)
Note: use immediately

Tris-glycine SDS-Polyacrylamide Gel (18% Separating)
30% Polyacrylamide60millilitre (ml)
1.5M Tris(pH8.8)25millilitre (ml)
Ammonium persulfate (10%)1millilitre (ml)
SDS (10%)1millilitre (ml)
TEMED0.04millilitre (ml)
dH2O13millilitre (ml)
Total volume100millilitre (ml)
Note: use immediately

Tris-glycine SDS-Polyacrylamide Gel (5% Stacking)
30% Polyacrylamide1.7millilitre (ml)
1M Tris(pH6.8)1.25millilitre (ml)
Ammonium persulfate (10%)0.1millilitre (ml)
SDS (10%)0.1millilitre (ml)
TEMED0.01millilitre (ml)
dH2O6.8millilitre (ml)
Total volume10millilitre (ml)
Note: use immediately

Tris-glycine SDS-Polyacrylamide Gel (8% Separating)
30% Polyacrylamide26.7millilitre (ml)
1.5M Tris(pH8.8)25millilitre (ml)
Ammonium persulfate (10%)1millilitre (ml)
SDS (10%)1millilitre (ml)
TEMED0.06millilitre (ml)
dH2O46.3millilitre (ml)
Total volume100millilitre (ml)
Note: use immediately

Tris-glycine SDS-Polyacrylamide Gel loading buffer (2X)
1M Tris-HCl (phH6.8)1.6millilitre (ml)
10% SDS4millilitre (ml)
Glycerol (100%)2millilitre (ml)
B-mercaptoethanol1millilitre (ml)
Bromophenol blue4milligram (mg)
ddH2O1.4millilitre (ml)
Total volume10millilitre (ml)
Note: store at 4 degree

Tris-glycine SDS-Polyacrylamide Gel loading buffer (3X)
1M Tris-HCl (phH6.8)2.4millilitre (ml)
20% SDS3millilitre (ml)
Glycerol (100%)3millilitre (ml)
B-mercaptoethanol1.6millilitre (ml)
Bromophenol blue6milligram (mg)
Total volume10millilitre (ml)
Note: store at 4 degree

Tris-glycine SDS-Polyacrylamide Gel running buffer (10X)
Tris base30.3gram (g)
Glycine144gram (g)
SDS10gram (g)
ddH2O to1litre (l)
Total volume1litre (l)

Trypsin/EDTA (0.025% Trypsin 0.01% EDTA)
Trypsin125milligram (mg)
EDTA50milligram (mg)
HBSS500millilitre (ml)
Total volume500millilitre (ml)
Note: filter sterilize, aliquot and store at -20 degree

Trypsin/EDTA (0.05% Trypsin 0.02% EDTA)
Trypsin250milligram (mg)
EDTA100milligram (mg)
HBSS500millilitre (ml)
Total volume500millilitre (ml)
Note: filter sterilize, aliquot and store at -20 degree

Trypsin/EDTA (0.25% Trypsin 0.1% EDTA)
Trypsin1250milligram (mg)
EDTA500milligram (mg)
HBSS500millilitre (ml)
Total volume500millilitre (ml)
Note: filter sterilize, aliquot and store at -20 degree

X-gal (20 mg/ml)
X-gal (5-bromo-4-chloro-3-indolyl b-D-galactopyranoside)0.2gram (g)
DMF (dimethylformamide)10millilitre (ml)
Total volume10millilitre (ml)
Note: aliquot and store protected from light at -20 degree

YEPD medium
YPD media1litre (l)
Total volume1litre (l)

YP medium
Bacto-yeast extract 10gram (g)
Bacto-peptone20gram (g)
ddH2O to900millilitre (ml)
Total volume900millilitre (ml)
Note: autoclave

YPD medium
Bacto-yeast extract 10gram (g)
Bacto-peptone20gram (g)
ddH2O to900millilitre (ml)
20% dextrose100millilitre (ml)
Total volume1litre (l)
Note: autoclave 20% dextrose separately from the rest, combine and pour plates.

YPD plates
Bacto-yeast extract 10gram (g)
Bacto-peptone20gram (g)
Bacto-agar20gram (g)
ddH2O to900millilitre (ml)
20% dextrose100millilitre (ml)
Total volume1litre (l)
Note: autoclave 20% dextrose separately from the rest, combine and pour plates.

YPG plates
Bacto-yeast extract 10gram (g)
Bacto-peptone20gram (g)
Bacto-agar20gram (g)
ddH2O to600millilitre (ml)
7.5% glycerol400millilitre (ml)
Total volume1litre (l)
Note: autoclave 7.5% glycerol separately from the rest, combine and pour plates.

YT medium (2X)
Bacto-tryptone 16gram (g)
Bacto-yeast extract 10gram (g)
NaCl5gram (g)
ddH2O to1litre (l)
Total volume1litre (l)
Note: autoclave to sterilize

YT plates
Bacto-tryptone 16gram (g)
Bacto-yeast extract 10gram (g)
Bacto-agar 16gram (g)
NaCl5gram (g)
ddH2O to1litre (l)
Total volume1litre (l)
Note: autoclave to sterilize, cool to 65 degree, add antibiotic if desired, and pour into sterile petri dishes

YT top soft agar
Bacto-tryptone 8gram (g)
Bacto-yeast extract 5gram (g)
Bacto-agar 3.5gram (g)
NaCl2.5gram (g)
ddH2O to0.5litre (l)
Total volume0.5litre (l)
Note: Heat to melt agar. Distribute 50 ml aliquots in 100 ml dilution bottles; autoclave to sterilize.

dNTP (10 mM)
dATP (100 mM)0.1millilitre (ml)
dCTP (100 mM)0.1millilitre (ml)
dGTP (100 mM)0.1millilitre (ml)
dTTP (100 mM)0.1millilitre (ml)
ddH2O0.6millilitre (ml)
Total volume1millilitre (ml)
Note: aliquot and store at -20 degree

rATP (adenosine triphosphate 100mM)
dipotassium ATP 619milligram (mg)
Sterile ddH2O to10millilitre (ml)
Total volume10millilitre (ml)