What will be the sizes of the 2 restriction fragments if NO insert is present in pUC19?

Essentials of Pharmacology for Health Professions
7th Edition
ISBN:9781305441620
Author:WOODROW
Publisher:WOODROW
Chapter5: Abbreviations And Systems Of Measurement
Section: Chapter Questions
Problem 7CRQ
icon
Related questions
icon
Concept explainers
Question

Knowing that you are using HindIII and EcoRI to cut your plasmids, and that those two enzymes cut within the MCS, use the map of pUC19 provided below to compute:

  1. What will be the sizes of the 2 restriction fragments if NO insert is present in pUC19?
  2. What will be the sizes of the 2 restriction fragments if the approximately 317 bp RT-PCR product (insert from WT satC dimer) was ligated successfully into the SmaI site?
  3. What will be the sizes of the 2 restriction fragments if TWO approximately 317 bp RT-PCR products (2 ligated inserts from WT satC dimer) were ligated into the SmaI site?
"spi, ri-ouei, raui, raeni, riiri,
mll, PpuMI, PshAI, Psil, PspOMI,
acll, SanDI(x), SexAI, Sfil, SgrAl,
rfl(x), Stul, Styl, Swal, Tlil,
Xhol
Drdl 91
BsmBI 51
BSMBI 2683
BSTAPI 179
Ndel 183
Eco01091 2674
Kasl - Narl - Sfol 235
AatlI - Zral 2617,
Bgll 245
FspI 256
Pvul 276
Pvull 306
of available from NEB
BcIVI 2542
Sspl 2501
Bmrl 364
Acll 2297
BoeAI 387
lacZa.
Apol - EcoRI 396
Banll - Sacl - EC053KI 402
Хmnl 2294
Ассб51- Крпl 408
Bcgl 2215
MCS
Aval - BsoBI - Smal -
Scal 2177
ТspMI - Хmal 412
BamHI 417
xbal 423
Pvul 2066
Accl - Hincll - Salı 429
Avall 2059
RSPMI - BfuAI 433
sbft 434
PUC19
2,686 bp
BsrDI 1935
Psti 435
Acll 1924
Sphi 441
Hindi 447
FspI 1919
Pvull 628
Avall 1837
NMEAIII 1822
Til 641
Bgll 1813
Врml 1784
BsaXI 659
BSPQI - Sapl 683
Tfil 781
BsrFI 1779
Bsal 1766
AflIII - Pell 806
BsrDI 1753
Drdl 908
Bmrl 1744
ori
Ahdl 1694
BcIVI 1015
BseYI 1110
AIWNI 1217
BceAI 1292
Fig. 1. Restriction map of pUC19 (2686 bp). Image obtained from the New England Biolabs e-catalog
(https://www.neb.com/products/n3041-puc19 vector#Product%20Information_Properties%20&%20Usage). Note
the presence of the lacZa gene, and note that the Smal site (nt 412) into which (ideally) your insert was
ligated is found within the lacZ ORF. Enzymes in the multiple cloning site (MCS) only cut the plasmid
once; enzymes not part of the MCS that are listed in boldface also only cut the plasmid once.
Transcribed Image Text:"spi, ri-ouei, raui, raeni, riiri, mll, PpuMI, PshAI, Psil, PspOMI, acll, SanDI(x), SexAI, Sfil, SgrAl, rfl(x), Stul, Styl, Swal, Tlil, Xhol Drdl 91 BsmBI 51 BSMBI 2683 BSTAPI 179 Ndel 183 Eco01091 2674 Kasl - Narl - Sfol 235 AatlI - Zral 2617, Bgll 245 FspI 256 Pvul 276 Pvull 306 of available from NEB BcIVI 2542 Sspl 2501 Bmrl 364 Acll 2297 BoeAI 387 lacZa. Apol - EcoRI 396 Banll - Sacl - EC053KI 402 Хmnl 2294 Ассб51- Крпl 408 Bcgl 2215 MCS Aval - BsoBI - Smal - Scal 2177 ТspMI - Хmal 412 BamHI 417 xbal 423 Pvul 2066 Accl - Hincll - Salı 429 Avall 2059 RSPMI - BfuAI 433 sbft 434 PUC19 2,686 bp BsrDI 1935 Psti 435 Acll 1924 Sphi 441 Hindi 447 FspI 1919 Pvull 628 Avall 1837 NMEAIII 1822 Til 641 Bgll 1813 Врml 1784 BsaXI 659 BSPQI - Sapl 683 Tfil 781 BsrFI 1779 Bsal 1766 AflIII - Pell 806 BsrDI 1753 Drdl 908 Bmrl 1744 ori Ahdl 1694 BcIVI 1015 BseYI 1110 AIWNI 1217 BceAI 1292 Fig. 1. Restriction map of pUC19 (2686 bp). Image obtained from the New England Biolabs e-catalog (https://www.neb.com/products/n3041-puc19 vector#Product%20Information_Properties%20&%20Usage). Note the presence of the lacZa gene, and note that the Smal site (nt 412) into which (ideally) your insert was ligated is found within the lacZ ORF. Enzymes in the multiple cloning site (MCS) only cut the plasmid once; enzymes not part of the MCS that are listed in boldface also only cut the plasmid once.
Expert Solution
trending now

Trending now

This is a popular solution!

steps

Step by step

Solved in 3 steps with 1 images

Blurred answer
Knowledge Booster
Bacterial genomics
Learn more about
Need a deep-dive on the concept behind this application? Look no further. Learn more about this topic, biology and related others by exploring similar questions and additional content below.
Similar questions
  • SEE MORE QUESTIONS
Recommended textbooks for you
Essentials of Pharmacology for Health Professions
Essentials of Pharmacology for Health Professions
Nursing
ISBN:
9781305441620
Author:
WOODROW
Publisher:
Cengage