MBLG1001 Calculations_1 Answers page 1

Calculations
(Try, wherever possible, to express your answer with numerical values between 0.1 and
1000. Change the prefix on the units rather than use scientific notation). Each question is
worth 2 marks. Please show all working, as part marks will be given for working even if
the final answer is incorrect. Submit your handwritten assignments at the beginning of the
next lab session to your demonstrator.

Question 1

Convert the following values to sensible numbers (between 0.1 and 1000) by changing the prefix:
(i) 0.09 L
(ii) 6,000 g
(iii) 0.007 g/100 mL
(iv) 2,000 nmol/min/mg protein

Answers

(i) 0.09 L = 90 mL
(ii) 6,000 g = 6 mg
(iii) 0.007 g/100 mL = 7 mg/100 mL
(iv) 2,000 nmol/min/mg protein = 2 mol/min/mg protein

Question 2
You have been provided with the following solutions of a compound (mol. wt. = 200). How many
moles would 100 l of each solution contain?
(i) 50 mM
50 moles/ml  5 moles in 100 l
(ii) 0.4 % (w/v)
0.4 g/100 ml  0.4 mg/ 100 l  0.4/200 = 2 moles.
(iii) 2 g/dL
2 g/dL = 2 g/100 ml = 2 mg/ 100 l  2000/200 = 10 moles
(iv) 16 mg/ml
1.6 mg/100 l  1600/200 = 8 moles.
MBLG1001 Calculations_1 Answers page 2

Question 3
You have a 5 mg/ml solution of an amino acid (mol. wt. = 125). You are required to add 0.4
moles to a test tube. How much would you pipette?
5/125 = 0.04 M = 40 mM  40 umoles/ml  40 nmoles/ul. We need 0.4 umoles OR 400 nmoles
 400/40 = 10 ul.

Question 4
You need a 10 mM solution of a potent (and dangerous) transcription inhibitor (mol. wt. 500) for
an experiment. To avoid handling the powder form of this compound you decide to make up the
whole bottle rather than attempt to weigh out a portion. The compound comes in 200 mg batches.
What volume would you add to one of these bottles to make up a 10 mM solution?

200/500 = 0.4 mmoles. 10 mmoles/L  10 umoles/ml. We need the volume for 400 umoles.
Volume = 400/10 = 40 ml.

Question 5
A reaction has been designed to be carried out in a 4 ml total volume. It requires a final
phenylalanine concentration of 5 mM. If the protocol allows 50 l for the phenylalanine addition,
what concentration would you make up the stock solution?
Final concentration is 5 mM  5 moles/ml  20 moles in 4 ml. You need 20 moles in 50 l
 20/50 = 0.4 moles/l  0.4 M.

Question 6
A particular cell line will grow well in a standard medium (DMEM) provided it is supplemented
with extra glutamine. The standard medium contains 1 mM glutamine but this cell line grows
better with 5 mM glutamine. How much more glutamine would you need to weigh out and add to
750 mL of the DMEM medium? The molecular weight of glutamine is 120.
750 mL of supplemented DMEM would contain 5*120 mg * 0.75 = 450 mg. The unsupplemented
DMEM would contain 90 mg of glutamine. You would need to add 450 – 90 = 360 mg. Equally
you could approach this by the following logic: You need to supplement the medium with 4 mM
glutamine so 0.75*4*120 mg = 360 mg.

Question 7
One of the model organisms used in molecular biology is the worm, Caenorhabditis elegans or C.
elegans for short. The medium used to grow these worms is quite complex. One of the components
of this medium is called S basal. It is composed of 0.1 M NaCl, 0.05 M potassium phosphate and
0.0005% (w/v) cholesterol. You have the following stock solutions and powders available: NaCl
powder (molecular weight 58.4), 1 M potassium phosphate, and a 5 mg/ml stock of cholesterol
(mol. wt. 386.7). How would you make up 250 ml of S basal?

Start with NaCl: 0.1 mole/l → 100 mmol/l → 25 mmol/250ml now 25*58.4=1460mg=1.46g NaCl
powder. Then 0.05 mmol/ml→12.5mmol/250ml and the stock is 1 mol/l→12.5 mmol/12.5 ml so
we need 12.5 ml 1 M potassium phosphate. Finally the cholesterol, 0.0005 g/100 ml→0.5
MBLG1001 Calculations_1 Answers page 3

mg/100 ml→1.25 mg/250ml and the stock is 5mg/ml→1.25mg/0.25ml so we need to add 250 ul
of stock cholesterol (don’t be distracted by the molecular weight). Then make up to 250 ml with
water.

Question 8
A citrate buffer must be made up to a final concentration of 250 mM. You need to make up 2 L.
You have two stock solutions from which to make this buffer; 0.5 M citric acid (C6H8O7) and a 0.5
M solution of the conjugate base, sodium citrate (C6H5O7Na3). You need to add 3 times as much of
the acid form than the base form to achieve the desired pH. How will you make up this solution?

The final conc is 0.25 M and the stock solutions are 0.5 M → 1 in 2 dilution → 1 l of the stock
solutions + 1 l of water. The 1 l has to be split 3 parts acid to 1 part base → 1000/4 = 250 ml/part.
Hence of the acid form we need 3*250 = 750 ml and of the base form 250 ml.

Question 9
Z buffer, a buffer used for growing up bacteria for cloning, contains 50 mM -mercaptoethanol
(mol. wt. 78.3), a reducing agent which stabilises many enzymes. -mercaptoethanol is a liquid
with a density of 1.12 g/mL. If you need to make up 200 mls of Z buffer how much -
mercaptoethanol would you need to add?
-mercaptoethanol = 1120 g/L  1120/78 M  14.35 M = 14.35 mmoles/ml. 200 mls of 50 mM
 50*200 umoles = 10 mmoles. How much do we need = 10/14.35 ml = 697 ul.

Question 10
A method requires 0.5 % (w/v) sodium acetate. You need to make up 500 ml but you only have the
hydrated form of Na Acetate available (CH3COONa.3H20 mol. wt. = 136). How much would you
need to weigh out?
You need 500 ml of 0.5 % NaAc  0.5*5 g = 2.5 g made up to 500 ml. However if you weigh out
the hydrated form you will underestimate it. Mol. wt. of NaAc = 136 - 3*18 = 82. You will need to
weigh out 2.5*136/82 = 4.15 g to have the same amount of NaAc.

Question 11
Nitric acid (HNO3) is supplied as a concentrated liquid. A bottle was assayed and found to have a
density of 1.42 g/ml and a purity of 70%. The mol. wt. of HNO3 is 63. What is the molarity of this
solution?
1.42 g/ml = 1420 g/L. 70% of 1420 = 994 g/63 = 15.8 M
MBLG1001 Calculations_1 Answers page 4

Question 12

If the average molecular weight of a deoxynucleotide monophosphate (dNMP), loosely termed a

base when describing genome sizes, is 330, hence a base pair is 660, how much genomic DNA
would a yeast cell contain? The haploid yeast genome contains 1.5 * 107 base pairs, although the
cell is diploid. How much DNA would 1 million (106) cells contain?

If the haploid yeast genome contains 1.5 * 107 bp the diploid will contain 3*107 bp. The molecular
weight of the yeast genome is 3*660*107 g/mole. 1 molecule would weigh
3*660/6.022 *107 *10-23 = 3.3*10-14 g = 33*10-15 g or 33 fg. If 1 yeast cell contains 33 fg then 1
million yeast cells would contain 33 ng DNA.