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Reto's Buffer & Media Book

Reto's Buffers & Media Book

These Buffers and Media are used in the Protocols of our Lab

Index:  A B C D E F G H I J K L M N O P Q R S T U V W X Y Z

08-15 Buffer
Compound
MW
Final concentration
200 ml
Glycine 75.07 
0.1 M
1.5 g
1 M MgCl2  
 10 mM
2 ml
KOH  
to pH 9
  
H2O     to 200 ml

A

Acetic Acid 1 M
Compound
MW
Final concentration
50 ml
Acetic Acid 98% (~17 M)
 
1 M
3 ml
H2O    
to 50 ml

Acrylamide/Bisacrylamide
Compound
MW
Final concentration
500 ml
Acrylamide
71.08
20 %
100 g
Bisacrylamide
154.17
0.8 %
4 g
H2O    
to 500 ml

Alkaline Lysis Buffer A
Compound
MW
Final concentration
500 ml
Glucose
180
50 mM
4.5 g
Tris
121
25 mM
1.5 g
EDTA 0.25 M  
10 mM
20 ml
H2O    
to 450 ml
Adjust pH 8.0 with HCl
H2O, Autoclave, store at 4°C    
to 500 ml

Alkaline Lysis Solution 1
Compound
MW
Final concentration
100 ml
Lysozyme  
0.2 mg/ml
0.02 g
Alkaline Lysis Buffer A    
to 100 ml

Alkaline Lysis Solution 2
Compound
MW
Final concentration
50 ml
SDS 20 %  
1 %
2.5 ml
NaOH 10 M  
0.2 M
1 ml
H2O    
to 50 ml

Alkaline Lysis Solution 3
Compound
MW
Final concentration
50 ml
KAc
98
3 M
14.7 g
Acetic Acid 100 %  
11.5 %
5.75 ml
H2O    
to 50 ml

Amido Black
Compound
MW
Final concentration
100 ml
Amidoblack
98
0.1 %
0.1 g
Acetic Acid 100 %  
7.5 %
7.5 ml
Methanol  
20 % (v/v)
20 ml
H2O    
to 100 ml

Aminoacid Concentration in Media
Constituent
Final mg/l
Stock
ml stock for 1 liter medium
Adenine sulfate
20
2 mg/ml
10
Uracil
20
2 mg/ml
10
L-tryptophan
20
 2 mg/ml
10
L-histidine-HCl
20
2 mg/ml
10
L-arginine-HCl
20
2 mg/ml
10
L-methionine
20
2 mg/ml
10
L-tyrosine
30
3 mg/ml
10
L-leucine
30
3 mg/ml
10
L-isoleucine
30
3 mg/ml
10
L-lysine-HCl
30
3 mg/ml
10
L-phenylalanine
50
5 mg/ml
10
L-glutamic acid
100
10 mg/ml
10
L-aspartic acid
100
10 mg/ml
10
L-valine
150
15 mg/ml
10
L-threonine
200
20 mg/ml
10
L-serine
400
40 mg/ml
10

3-Aminophtalhydrazide 250 mM
Compound
MW
Final concentration
10 ml
3-Aminophtalhydrazide
177
250 mM
442 mg
H2O    
to 10 ml

Ammoniummolybdate 4.2 %
Compound
MW
Final concentration
50 ml
Ammoniummolybdate x 4H2O
236
4.2 %
2.1 g
H2O    
to 30 ml
HCl fuming 37 % (~12 M)  
4 M
16.7 ml
H2O    
to 50 ml

Ampicillin 20 mg/ml pH 8.0
Compound
MW
Final concentration
15 ml
Ampicillin
349.4
20 mg/ml
300 mg
H2O    
to 10 ml
Adjust pH 8.0 with NaOH
H2O, Filter sterilize    
to 15 ml

Antibiotics
Antibiotics
mg/ml on plates
Action
 Resistance Mechanism
Ampicillin
50
 Inhibits cell wall synthesis by inhibiting formation of the peptidoglycan crosslink b-lactamase (286 aa) is secreted into the periplasm and cleaves lactam antibiotic
Chloramphenicol
20
 Inhibits protein synthesis by interacting with the 50S ribosomal subunit and inhibiting the peptidyltransferase reaction. 216 aa protein cytoplasmic protein which acetylates chloramphenicol
Kanamycin 
Neomycin
30
 Inhibits translocation and elicits miscoding by binding to the 30S ribosomal subunit. 256 aa aminoglycoside phosphotransferase phosphorylates kanamycin, neomycin, etc.
Tetracycline
12
 Inhibits protein synthesis by preventing binding of aminoacyl tRNA to the ribosome A site. 428 aa membrane protein catalyzing efflux of tetracyclin, probably by a cation symport mechanism.

APS
Compound
MW
Final concentration
10 ml
Ammoniumpersulfate
228.2
10 %
1 g
H2O, store at 4°C    
to 10 ml

Ascorbate 0.5 M pH 6.0
Compound
MW
Final concentration
50 ml
L-Ascorbic acid
176.1
0.5 M
4.4 g
H2O    
to 22 ml
Adjust pH 6.0 with NaOH 1 M
H2O    
to 50 ml

Asolectin (fatty acid free) 50 mg/ml
Compound
MW
Final concentration
50 ml
Asolectin (aceton treated)
 
50 mg/ml
2.5 g
Diethylether, store at -20°C    
to 50 ml

ATP 200 mM
Compound
MW
Final concentration
10 ml
Adenosine-5'-Triphosphate
551.1
200 mM
1.102 g
H2O    
to 5 ml
Adjust pH 6.5 with NaOH 1 M
H2O, store at -20°C    
to 10 ml

ATP Regenerating System (ARS) 40x
Compound
MW
Final concentration
1 ml
Phospoenolpyruvate (PEP)
208.0
20 mM
4.16 mg
Pyruvate Kinase (PK) 5 U/ml  
0.1 U/ml
20 ml
H2O, store at 4°C    
to 1 ml

B

BALP Protease Inhibitor Mix 21x
Compound
MW
Final concentration
10 ml
Aprotinin 1.5 mg/ml  
0.486 g/l
3.24 ml
PepstatinA 1 mg/ml in DMSO
686
0.131 g/l
1.3 ml
Leupeptin x 1/2H2SO4 10.5 mM
476
0.04 mM
37.5 ml
4-Aminobenzamidine x 2HCl
208
190 mM
396 mg
H2O    
to 10 ml

Bis-Tris 1M pH7.0
Compound
MW
Final concentration
100 ml
Bis-Tris
209.2
1 M
20.9 g
H2O    
to 50 ml
Adjust pH 7.0 with HCl      
H2O    
to 100 ml

Brij/Doc pH 8.0
Compound
MW
Final concentration
100 ml
EDTA 0.25 M
 
1 mM
0.4 ml
Tris 1 M pH 8.0  
10 mM
1 ml
Na-Deoxycholate 8 %  
0.4 %
5 ml
Brij 58  
1 %
1 g
H2O    
to 90 ml
Dissolve at 37°C, adjust pH 8.0
H2O, Filter sterilize     to 100 ml

BS3 Buffer pH7.5
Compound
MW
Final concentration
1000 ml
HEPES
238.3
100 mM
23.83 g
Dodecylmaltoside
 510.6
0.05 %
0.5 g
EDTA 0.25 M  
1 mM
4 ml
DTT
154
10 mM
1.54 g
H2O    
to 900 ml
Adjust pH 7.5 with NaOH 10M
H2O     to 1000 ml

C

CdSO4 1 M
Compound
MW
Final concentration
1.5 ml
CdSO4
208.5
1 M
313 mg
H2O    
to 1.5 ml

Chloroform
Compound
MW
Final concentration
50 ml
Chloroform  
 
48 ml
Isoamyl alcohol  
4 %
2 ml

CIP Buffer 4x
Compound
MW
Final concentration
15 ml
EDTA 0.25 M pH 8.0
 
200 mM
12 ml
Tris 1 M pH 8.0  
0.4 mM
6 ml
H2O    
3 ml

Citrate 1M pH5.O
Compound
MW
Final concentration
50 ml
Citrate x H2O
210
1 M
10.5 g
H2O    
to 35 ml
Adjust pH 5.0 with NaOH 10M
H2O     to 50 ml

Colony PCR Buffer 10x
Compound
MW
Final concentration
15 ml
Tris 1M pH6.8
 
125 mM
1.875 ml
KCl
74.5
0.5625 M
0.629 g
H2O, Autoclave    
to 15 ml

Coomassie Blue Solution
Compound
MW
Final concentration
1000 ml
Coomassie Blue (Serva Blau)
 
0.2 %
2 g
Acetic acid  
7.5 %
75 ml
Methanol  
50 %
500 ml
H2O    
to 1000 ml

Coumaric Acid 40 mM
Compound
MW
Final concentration
10 ml
p-Coumaric acid
164
40 mM
66 mg
H2O    
to 10 ml

CS/PBS
Compound
MW
Final concentration
200 ml
PBS, 20x
 
1x
10 ml
Newborn calf serum  
10 %
20 ml
H2O, Store frozen     200 ml

CuSO4 1 M
Compound
MW
Final concentration
25 ml
CuSO4
159.6
1 M
4 g
H2O    
to 25 ml

D

DEPC treated H2O



&
Compound
MW
Final concentration
1000 ml
H2O