Enzyme(s) used PstI ECORI HincII Band sizes observed (kb) 5.3, 2.7 5.4, 2.6 Smal Xbal ВатHI 4.5, 3.5 6.7,1.3 4.0 (high intensity band) 3.9, 3.7,0.4

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Restriction mapping sample question
You have a 5.3 kb PstI fragment cloned into the PstI
site of the vector pUC19, which is 2.7 kb in size. This
vector has unique sites for the following enzymes in a
multiple cloning site:
PstI, HincII, Xbal, BamHI, SmaI, EcoRI
A restriction map of the 5.3 kb insert is prepared. The
recombinant plasmid is digested with the enzymes
listed above in single digests, and then several
combinations of enzymes are tested in double
digests. The following bands are observed when the
digests are run on a gel:
Enzyme(s) used
PstI
ECORI
HincII
Band sizes observed (kb)
5.3, 2.7
5.4, 2.6
4.5, 3.5
6.7, 1.3
| 4.0 (high intensity band)
3.9, 3.7, 0.4
4.0, 3.5, 0.5
3.5, 2.6, 1.9
3.7, 3.6, 0.4, 0.3
3.7, 2.2, 1.7, 0.4
3.7, 3.0, 0.9, 0.4
3.9, 3.5, 0.4, 0.2
Smal
Xbal
ВатHI
HinclI + Xbal
HincII + ECORI
XbaI + BamHI
ECORI + BamHI
Smal + BamHI
HincII + BamHI
Use the data above to construct a map of the cloned
insert. Note that fragments smaller than 100 bp will
not usually be visible on a gel, and that 2 fragments
with a similar size will run at the same spot on a gel –
resulting in a brighter band.
Transcribed Image Text:Restriction mapping sample question You have a 5.3 kb PstI fragment cloned into the PstI site of the vector pUC19, which is 2.7 kb in size. This vector has unique sites for the following enzymes in a multiple cloning site: PstI, HincII, Xbal, BamHI, SmaI, EcoRI A restriction map of the 5.3 kb insert is prepared. The recombinant plasmid is digested with the enzymes listed above in single digests, and then several combinations of enzymes are tested in double digests. The following bands are observed when the digests are run on a gel: Enzyme(s) used PstI ECORI HincII Band sizes observed (kb) 5.3, 2.7 5.4, 2.6 4.5, 3.5 6.7, 1.3 | 4.0 (high intensity band) 3.9, 3.7, 0.4 4.0, 3.5, 0.5 3.5, 2.6, 1.9 3.7, 3.6, 0.4, 0.3 3.7, 2.2, 1.7, 0.4 3.7, 3.0, 0.9, 0.4 3.9, 3.5, 0.4, 0.2 Smal Xbal ВатHI HinclI + Xbal HincII + ECORI XbaI + BamHI ECORI + BamHI Smal + BamHI HincII + BamHI Use the data above to construct a map of the cloned insert. Note that fragments smaller than 100 bp will not usually be visible on a gel, and that 2 fragments with a similar size will run at the same spot on a gel – resulting in a brighter band.
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