LAB PREP NOTEBOOK
CCC – M. Lawliss
Table of contents:
Buffers:
- Catalase Lab Buffer (100mM Phosphate
Buffer)
- TBE Buffer
-TAE Buffer
Dilutions:
- H2O2 dilutions for Catalase
- Hydrochloric Acid
Standard Solutions:
- 600 mM Sucrose
- 0.15 Cobalt Chloride
- 100mM Glucose
- 20mM NaCl
- Chloride Ion Standards for Env Sci
-0.05M Conductivity Solutions
-10 mM Caffeine
-Cal. Sol. for Conductivity Meter Env-Sci
Bio 101 salt and glucose solutions gradient
Other:
- Fake Blood
- Potato Extract
- Benedict’s Solution
- Fake Urines
- I2 Clock
- Methoxide for BioFuels Lab
Demo preps:
- Stoplight Reaction
100mM Phosphate Buffer
• Dissolve the following then dilute to 1.0 L total
volume:
– 8.702g Potassium Phosphate Dibasic
– 6.805g Potassium Phosphate Dibasic
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TBE Buffer
• Makes 1L of 10x concentration:
– 108g Tris (hydroxymethyl)-aminomethane
C4H11NO3 [121.14 g/mol]
– 55g Boric Acid H3BO3 [61.84 g/mol]
– 20ml of 0.5M EDTA (Ethylenediamenetetraacetic
acid Disodium salt) or 7.5g of EDTA –Disodium Salt
[372.2 g/mol]
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TAE Buffer
• Makes 1L of 10x concentration:
– 108g Tris (hydroxymethyl)-aminomethane
C4H11NO3 [121.14 g/mol]
– 10.9g Glacial Acetic Acid [60.05 g/mol]
– 20ml of 0.5M EDTA (Ethylenediamenetetraacetic
acid Disodium salt) or 7.5g of EDTA –Disodium Salt
[372.2 g/mol]
– Adjust pH to 8.5 with KOH
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600mM Sucrose
• For a 200ml volume. Dissolve the following
then dilute to 200ml total volume:
– 41.076g Sucrose
200ml | 0.60 mole_| 342.30g Sucrose_| = 41.047g sucrose
| 1000ml
| 1 mole Sucrose |
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0.15 M CoCl2*6H2O
• To make 500ml
500ml | 0.15 mole_| 237.93g CoCl2_| = 17.845g CoCl2
| 1000ml | 1 mole CoCl2 |
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100mM Glucose
• For a 250ml volume. Dissolve the following
then dilute to 250ml total volume:
– 4.504g Glucose
250ml | 0.100 mole_| 180.16g Glucose_| = 4.504g Glucose
| 1000ml
| 1 mole Glucose |
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20mM NaCl
• For a 250ml volume. Dissolve the following
then dilute to 250ml total volume:
– 1.292g NaCl
250ml | 0.020 mole| 58.44g NaCl__| = 0.292g NaCl
| 1000ml
| 1 mole NaCl |
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Cl-1 Ion Solution Prep
Mass Chlorine
10mg/L
100mg/L
300mg/L
10mg
100mg
300mg
164.7mg/L
494.2mg/L
0.1647g NaCl
0.4942g NaCl
Mass NaCl/Liter 16.47mg/L
For 1 Liter of
solution 0.01647g NaCl
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Conductivity Standards 0.05M
Substance
Formula Mass
Amount to use
Dissolve and
Dilute to:
CaCl2*2H2O
147.02g/mol
3.6755g
500ml
AlCl3*6H2O
241.43g/mol
NaCl
58.44g/mol
H3BO3
61.83g/mol
C2H6O2
62.07g/mol
6.0358g
1.461g
1.546g
1.5518g
500ml
500ml
500ml
500ml
(ethylene Glycol)
CH3OH
32.04g/mol
Dens = 0.7910 g/ml
0.801g ->
1.01ml
500ml
Acetic Acid
(17.4M)(vol)=(1000ml)(0.05M)
HCl
(12.1M)(vol)=(1000ml)(0.05M)
2.87ml
4.13ml
1000ml
1000ml
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10mM Caffeine
• 0.388g caffeine in 200ml
200ml | 0.010 mole | 194.19 g Caff__| = 0.292g Caffeine ~0.15%
| 1000ml | 1 mole Caff
|
Tobacco Tea –
(approx. 0.011 mg/ml) ~0.001%
2 regular cigarettes’ worth of tobacco in 500ml hot distilled
water. Filter.
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Sodium Methoxide
• Ratio 0.5g NaOH in 15ml of MeOH
• 8.333g NaOH in 250ml MeOH
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Calibration Solution for Conductivity Meter (0.01
M KCl, 1411 μS at 25°C)
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a) Place 2-3 g of AR potassium chloride (KCl) in a 50 ml beaker and dry in an oven for 3-5 hours at 105°C
then cool to room temperature in a desiccator.
b) Weigh 0.746 g of KCl into another 50 ml beaker.
c) Dissolve the KCl in the beaker with good quality de-ionised water, e.g. ‘Option water’.
d) Pour the solution from the 50 ml beaker into a 1.0 litre volumetric flask. Thoroughly rinse the beaker
into the flask with de-ionised water. Fill the flask almost to the mark then shake it to fully dissolve the
KCl. Top up to the mark and invert the flask a few more times so the solution is fully mixed.
e) Rinse out the storage bottle with de-ionised water then shake dry and rinse again with a small amount
of the KCl solution then shake dry again.
f) Fill the storage bottle with the new solution and label with appropriate information including the expiry
date. This solution has a shelf life of 1 year.
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Dilutions of 3% H2O2
Desired % conc.
QTY of 3% to use (ml)
Dilute to:
0.3%
50.0
500ml
0.6%
100
500ml
0.8%
133
500ml
1.0%
167
500ml
1.5%
250
500ml
2.0%
333
500ml
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Hydrochloric Acid Dilutions
• From 12.1M HCl stock (37%):
To make 1 L of
n concentration
Math
Use x ml of
12.1M HCl
Dilute to:
6 Molar HCl
(12.1M)(vol)=(1000ml)(6M)
495.9 ml
1.0 Liter
5 Molar HCl
(12.1M)(vol)=(1000ml)(5M)
413.2 ml
1.0 Liter
2 Molar HCl
(12.1M)(vol)=(1000ml)(2M)
165.3 ml
1.0 Liter
0.2% HCl
(37%)(vol)=(1000ml)(0.2%)
5.4 ml
1.0 Liter
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Bio 101 salt and glucose solutions gradient
Volume of solution
Molarity
200 ml
of
Solution
0.050
0.080
0.150
0.300
1.000
0.5844
0.93504
1.7532
3.5064
11.688
0.29%
0.47%
0.88%
1.75%
5.84%
1.8016
2.88256
5.4048
10.8096
36.032
0.90%
1.44%
2.70%
5.40%
18.02%
Form Weight
NaCl
58.44
grams of solute
%
Molec Weight
Glucose
(anhyd)
180.16
grams of solute
%
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Fake Blood Recipe
To make 1 Liter of Fake
Blood
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–
–
–
–
225 ml corn syrup
90 ml Sobe citrus drink
3 TBS choc powder
15 drops red dye
3 TBS corn starch
In a second mug place 1 level tsp of corn starch
Add 1-2 tsp of water and stir until it is
completely mixed and will flow.
In a coffee mug boil 100ml of water (~3.5 oz)
Dump the boiling water into the mug with the
corn starch, stir to mix this completely.
Allow to cool until warm.
Add ½ tsp baking cocoa.
Add 12 drops of red food coloring
When completely cooled, add 3-4 squirts of
hand sanitizer
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Potato Extract for Catalase
• This ratio is to standardize the prep only.
– 250g of potato
– 100ml of 100mM Phosphate Buffer
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Benedict’s Solution
1 L of Benedict's reagent can be prepared from:
- 100 g of anhydrous sodium carbonate,
- 173 g of sodium citrate and
- 17.3 g of copper(II) sulfate pentahydrate
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• Normal Urine (2L):
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–
–
–
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Fake Urines
Mix 36.4 grams urea with 1.5L distilled water until completely dissolved
Add: 15g NaCl; 9g KCl; 9.6g Na3PO4
Adjust pH to range between 5- 7 using HCl and NaOH
Dilute with water until Spec Grav is between 1.015 and 1.025
Add 0.1g Albumin (bovine) and 4g of creatine
Conditions simulated ( per 1L of normal urine from above):
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Glycosuria: Add 2.5 – 5g glucose
Proteinuria: Add 1g Albumine
Ketonuria: Add 0.1g Acetoacetic acid OR 1 ml acetone
Low pH: add HCl until pH is 4.0 – 4.5
High pH: add NaOH until pH is between 8.0 – 9.0
Hyposenthuria: Add D-H2O until Spec Grav is below 1.010
Hematuria: Add 0.5g bovine hemoglobin
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STOPLIGHT Reaction
• 300ml total volume, equal amounts of
– 150 ml of 0.133M Dextrose
– 150 ml of 1.0M NaOH
– Small amt of Indigo Carmine
150ml | 0.133 mole_| 180g Dextrose _| = 3.6g Dextrose
| 1000ml | 1 mole Dextrose |
150ml | 1.0 mole_| 40g NaOH _| = 6.0g NaOH
| 1000ml | 1 mole NaOH |
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I2 Clock
NaI
Sodium Iodide
0.2M
NaCl
Sodium Chloride
0.2M
Na2S2O3
Sodium Thiosulfate
0.01M
K2SO4
K2S2O8
Potassium Sulfate
Potassium Persulfate
0.2M
0.2M
* 2% Starch
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