1. You have the plasmid pUC18/19, which is a circular plasmid that consists of 2686 bp. What would the number of and length of the fragments be if you cut the plasmid with the following restriction enzymes or combination of enzymes? Give a schematic representation of the digestions. a. Pscl& Gsul b. Scal, Pdml & BsaXI c. Scal, Sspl. & Ebel

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Chapter2: Life's Chemical Basis
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Eco01091 2674 Pfol 46 BstAPI 179
Aatil 2617
Ndel 183
Sspl 2501
Ehel 235
Pdml 2204
Bogl 2215
Scal 2172
146/
lacz
406
MCS
236
409
PUC18/19
2686 bp
Bsax 659
Sapl 683
852
LAlI, Pscl 806
Gsul 1784
1626
Cfr101 1779
Eco311 1766
Eam11051 1694/
rep (PMB1)
\Cail 1217
\BseYl 1110
29
ATC COC GO0 TAC CA CT CAT G or E IST E CG
CA ATT Oir M
y e am m ens ccs e NG stt CGA ACG IAC GA CT CCA GCT GAG ATC TOC TAG GOG CCC RG CT CA GT TM GA an co a e A ec y
Lael-WWa las A Lan Sr O a in A O N Sr Sr W Ne
1. You have the plasmid pUC18/19, which is a circular plasmid that consists of 2686 bp.
What would the number of and length of the fragments be if you cut the plasmid with
the following restriction enzymes or combination of enzymes? Give a schematic
representation of the digestions.
a. Pscl & Gsul
b. Scal, Pdml & BsaXI
c. Scal, Sspl & Ehel
2. You have determined that the amplicon you want to clone has enzyme restriction sites
for Hindll, and ĘCORI, After investigation you have seen that the PUC18/19 plasmid
6
also have these enzyme restriction sites in its multiple cloning site (MCS on map
above). After enzyme digestion your amplicon is 854 bp long.
a. What length will the recombinant plasmid be after you have inserted your
amplicon? Show your calculation.
b. In the amplicon insert you have an enzyme restriction site for Ndel at 500 bp.
If you digest the recombinant plasmid with this enzyme what length will the
fragments be?
Transcribed Image Text:Eco01091 2674 Pfol 46 BstAPI 179 Aatil 2617 Ndel 183 Sspl 2501 Ehel 235 Pdml 2204 Bogl 2215 Scal 2172 146/ lacz 406 MCS 236 409 PUC18/19 2686 bp Bsax 659 Sapl 683 852 LAlI, Pscl 806 Gsul 1784 1626 Cfr101 1779 Eco311 1766 Eam11051 1694/ rep (PMB1) \Cail 1217 \BseYl 1110 29 ATC COC GO0 TAC CA CT CAT G or E IST E CG CA ATT Oir M y e am m ens ccs e NG stt CGA ACG IAC GA CT CCA GCT GAG ATC TOC TAG GOG CCC RG CT CA GT TM GA an co a e A ec y Lael-WWa las A Lan Sr O a in A O N Sr Sr W Ne 1. You have the plasmid pUC18/19, which is a circular plasmid that consists of 2686 bp. What would the number of and length of the fragments be if you cut the plasmid with the following restriction enzymes or combination of enzymes? Give a schematic representation of the digestions. a. Pscl & Gsul b. Scal, Pdml & BsaXI c. Scal, Sspl & Ehel 2. You have determined that the amplicon you want to clone has enzyme restriction sites for Hindll, and ĘCORI, After investigation you have seen that the PUC18/19 plasmid 6 also have these enzyme restriction sites in its multiple cloning site (MCS on map above). After enzyme digestion your amplicon is 854 bp long. a. What length will the recombinant plasmid be after you have inserted your amplicon? Show your calculation. b. In the amplicon insert you have an enzyme restriction site for Ndel at 500 bp. If you digest the recombinant plasmid with this enzyme what length will the fragments be?
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