Figure 2-5

DNA Replication
Fragment of DNA
(4 linked nucleotides)

Phosphodiester bond links

the nucleotides

3’ of one nucleotide to 5’
of the following nucleotide

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Figure 4.7 (Campbell, et al.)

DNA molecule

Proteins

Chromatin
fiber Chromosome

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Figure 10.7
Origin of Origin of
replication replication
Parental strands

Origin of
replication
Parental strand
Daughter strand

Bubble

Two daughter DNA molecules

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Figure 2-6

Each ‘old’ strand of DNA is used

as a template for a ‘new’ strand of
DNA

Semi-conservative replication

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Figure 2-10

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Figure 2-8

The ‘new’ strand is made 5’ 3’, while ‘old’ strand is

read 3’ 5’

Synthesis reaction is catalyzed by DNA polymerase

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 PCR: Artificial DNA Replication

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Unnumbered Figure 2-1

Central Dogma of Molecular Biology

(mRNA)

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Figure 2-17

Transcription: RNA production

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 Sense and Antisense;
Codons and Anti-codons

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 Sense Strand
(aka Coding Strand)
(aka Non-template Strand)

Anti-sense Strand
(aka non-coding Strand)
(aka Template Strand)

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 Sense Strand
(aka Coding Strand)
(aka Non-template Strand)

• If you search the sequence of a gene, you will be given

the information of the sense strand

• Eg. Human Insulin

-https://www.ncbi.nlm.nih.gov/nuccore/AH002844.2

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Figure 2-14

Transfer RNA (tRNA) molecules are the

‘adaptors’ for protein synthesis

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Figure 2-15

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Figure 2-13

Ribosomes in rough ER

Site of protein synthesis

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Figure 2-16

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Figure 2-18

RNA is synthesized in the

nucleus

It then exits the nucleus to the

cytoplasm

Available to ribosomes in rough

ER for protein synthesis

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Table 2-3

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 Molecular Biotechnology Tool
• Determine concentration of live bacteria/cells

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 Bacteria concentration
• OD600 of 1.0 = 8 x 108 cells/ml
- doesn’t distinguish between live and dead cells

• Pfu

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 Cell counting
• Haemocytometer (with trypan blue)

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 Cell counting
• Haemocytometer (with trypan blue)

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 Cell counting
• Haemocytometer (with trypan blue)

Live cells “spit out” trypan blue; dead ones cannot

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 Cell counting
• Haemocytometer (with trypan blue)

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 Cell counting

Dimensions

=1 mm x 1 mm x 0.1 mm
=0.1 cm x 0.1 cm x 0.01 cm
=0.0001 cm3
=10-4 cm3
=10-4 ml

Coverslip designed to give height of 0.1 mm

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 Cell counting

25
=25 cells in 10-4 ml

Therefore
=25 x 104 cells/ml

=2.5 x 105 cells/ml

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 Cell counting
Average = (35 +25+ 28 + 32)
4
35 25
=30 cells in 10-4 ml

Therefore
=30 x 104 cells/ml
28 32
=3.0 x 105 cells/ml

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 Cell counting

250 240
Can dilute to manageable
numbers

Eg. Dilute 10 fold

260 270

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 Cell counting
Average = (25 +24+ 26 + 27)
4

=25.5 cells in 10-4 ml

25 24
Therefore
=25.5 x 104 cells/ml

=2.55 x 105 cells/ml

26 27 Must account for dilution

x 10 (dilution factor)

=2.55 x 106 cells /ml

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 Clicker counter

250 240

260 270

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 Automated Haemocytometer

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