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Color Plate 1 Color Plate 4

Color Plate 2 Color Plate 5

Color Plate 3 Color Plate 6

Color Plate 7 Color Plate 1 0

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Color Plate 8
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Color Plate 11

Color Plate 9 Color Plate 1 2

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Color Plate 13 Color Plate 16

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Color Plate 14 Color Plate 17

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Color Plate 15
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Color Plate 18
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Color Plate 19 Color Plate 20

Immunoglobulin
Molecule
Dimeric IgA Molecule
Fab Fab

Fab Fab
Color Plate 21 Color Plate 22

Tube N o . 1 2 3 4 5 6 7 8 9 1 0 1 1
" N / ^ ' X / ^ ' N / ' ' " ^ / ' *

Agglutination 0 1+ 2 + 4 + 4 + 3 + 3 + 2 + 1 + 1 + 0
Color Plate 23
 Tube No. 1 2 3 4 5 6 7 8 9 1 0

Agglutination Pos Pos Pos Pos Pos Pos Pos Pos Neg Neg
Color Plate 24 Color Plate 25

Color Plate 26 Color Plate 27

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Color Plate 28 Color Plate 29

Color Plate 30 Color Plate 31

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Color Plate 32
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Color Plate 35

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Color Plate 34 Color Plate 37
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Color Plate 38 Color Plate 41

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Color Plate 39
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Color Plate 42

Color Plate 40 Color Plate 43

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Color Plate 45 Color Plate 48

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Color Plate 46 Color Plate 49

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Color Plate 50 Color Plate 53

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Color Plate 54

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Color Plate 52 Color Plate 55
 M B D
Digest DNA with restriction enzyme
Separate by electrophoresis
5000 1 2 3 4 5
4000
3000 BH •• • E=I C=3
2000 23kb ••

1000 9.4 mm

6.6
900
—
800 4.1
—
700 I I 2.3
2.1 -
1 1
600
0.5
—
500 Chemical depurination
Denaturation
400
Neutralization
Transfer to membrane
Color Plate 56 Bind DNA to membrane

Hybridize membrane with 32P labeled probe

Wash off excess probe
Expose to X-ray film
Develop autoradiogram
1 2 3 4 5
23kb—
9.
6.
4.1
2.
2.1—

0.5—

Color Plate 57
Cycle 1: Start with (1) Double-stranded DNA TEMPLATE ycle 2: Start with 2 double-stranded cDNA intermediates containing target sequence

' - G G C A A T A T T G C G A A G C C T G T C T - 3 '

Step 1: Denaturation

1A 5' -G G C A A T A ss TARGET C C T G T C T - 3'

Step 1: Denaturation to 4 ssDNA templates
-C C G T T
1A 5'-G G C A A ss TARGET C T G T C T- 3'
Step 2: Annealing of primers
SS TARGET
Reverse primer 5 ' G A
G C A A T ss TARGET
-G G C A A T A ss TARGET C C T G T C T- 3' ss TARGET
Step 2: Annealing of primers
ss Step 3: Extension of primers to form 4 double-stranded copies
Step 3: Extension of primers

I
dNTPs:| 2 1 E C S • Buffer: 1A 5'-G G C A A ds TARGET C T G T C T- 3'
3B 3'-C C G T T ds TARGET G [A C A
KC1,
IIIIIIIIIIIG G G Mg++, PH 8.3 2B C C G T T A T A A C G C T T C G G [A C A
4A G C A A T A T T G C G A A G C C T G T
1A 5' - G G C A A T A T T G C G A A C G T C T- 3'
Precise length products become preferred templates (i.e., 4A & 4B)
a
4B C G T
2A G C A A T A T T G C G A A G C C T G T C T - 3 '

3A G C A A T ds TARGET C T G T C T-

Cycle 3 to 30: Repeat 3 step cycle.

End of cycle 1: 2 double stranded DNA products At the end of 30 cycles = 23( copies of target sequence
Color Plate 58a
Setc.A 5' G C A A T A T T G C G A A G C C T G T C
Setc.B 3'

Note: Precise length products are bounded by primer sequences

Color Plate 58b
 M 1 2 3 4 5 + Neg

500 bp
250 bp

Color Plate 59

22 25

600

Color Plate 60