pUC 18

1
Rational behind Part A
pUC 18 3 kb
Sal I
Sac I
Xba I
Hind III
Bam HI
Bam HI
Not drawn to scale
1.5 Kb BamHI fragment Inserted Creating ICE-1
molecular size 4.5 Kb
2
Use guide lines or graph paper
A) Scaled Map 1.5 Kb cDNA of ice fish
1,500
0
500
1,000
Mark on known vector sites
Xba I
Hind III
Sac I
Sal I
Bam HI - 3000, 1500
Bam HI
Bam HI
Sac I - 3500, 1000
Sac I
Sac I
Xba I - 3750, 750
Xba I
Xba I
Sal I - 3600, 900
Sal I
Sal I
Sac I / Sal I - 3000, 600 500, 400
Sal I
Sac I
Sal I
Sac I
200
400
400
Hind III - 3500, 400, 200 (sum 4100, either
second 400 or 2 more 200. As both double digests
have 400 band in addition to 300 and 100) Bands
therefore must be two 400)
Either
Hind III
Hind III
Hind III
Hind III
400
200
400
or
Hind III
Hind III
Hind III
Hind III
200
400
400
or
Hind III
SaI I (double digest 3500, 400, 300 200, 100)
Sac I (double digest 3000, 500, 400, 300 200,
100)
Sal I
Xba I
Hind III
Sac I
Sac I
Xba I
Sal I
Composite
3
Rational behind Part B
pUC 18 3 kb
Sal I
Sac I
Sph I
Pst I
Pst I
Not drawn to scale
7 Kb Pst I fragment of genomic DNA
Sac I / Sal I - 3000, 600 500, 400
Sal I
Sac I
Sal I
Sac I
Sac I / Sal I 400 bp fragment
4
0
5 Kb
10 Kb
B) Draw a scaled Map of the genomic clone
3 Kb vector
7 Kb Insert
Mark on known vector sites
Sph I
Sal I
Sac I
Sph I - 10000 labeled 10000
Sph I
labelled
Pst I -7000, 3000 labeled 7000
Pst I
Pst I
labelled
Sac I - 6000, 4000 labeled 6000
or
labelled
Sac I
Sac I
Either
Sac I
labelled
Sac I / Sph I - 6000, 300, 1000 labeled 6000
labelled
Sac I
Sac I
Sph I
labelled
SaI I - 5500, 3500, 1000 labeled 5000, 1000
labelled
labelled
Sal I
labelled
labelled
400 bp Sac I / Sal I fragment know taking up 2.5
Kb indicating the insertion of one or more intron
with combined molecular weight of 2.1 Kb.
Sac I/ SaI I - 4000, 3500, 1000, 1500 labeled
1500, 1000
Sac I
Sac I
Pst I
labelled
Pst I
Composite Map
Sac I
Sac I