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A commonly used plasmid that we will use in this lab is PUC19: (New England Biolabs) Indl 91 BamBI 51 Bsnlil 2683 Eco01091 2674 BstAPI 179 Ndel 183 Kasl - Narl - Sfol 235 Aatll - Zral 2617. Bell 245 Vpl 256 Prul 276 Pull 306 BiVI 2542. Sspl 2501 Hmrl 364 Aell 2297 Bcetl 387 Apel - Ecoti 396 Banll - Sacl - EcosKI 402 lecs1 - Kpnl 408 Xmnl 2294 laczu Begl 2215 MCS wal - BoBI - Smal- Scal 2177 TspMI - Xmal 412 Hamill 417 thal 23 Prul 2066 Aecl - linell - Sall 429 Avall 2059 PUC19 2,686 bp Sh 434 Pstl 35 BaDI 1935 Acll 1924 Fipl 1919 Sphl 441 Hindl 417 Prull 628 wall 1837 NmeI 1822 Bell 1813 Bpml 1784 Bsrfl 1779 Bsal 1766 Til 641 BsaXI 659 Bspol - Sapl 683 Thl 781 BarD 1753 Bnrl 1744 ANIII - Peil 006 Drdl 908 ori Ahdl 1694 BeiVI 1015 Bsell 1110 AlwNI 1217 kell 1292 A. Which plasmid is used? B. Which antibiotic was used?

A commonly used plasmid that we will use in this lab is PUC19: (New England Biolabs) Indl 91 BamBI 51 Bsnlil 2683 Eco01091 2674 BstAPI 179 Ndel 183 Kasl - Narl - Sfol 235 Aatll - Zral 2617. Bell 245 Vpl 256 Prul 276 Pull 306 BiVI 2542. Sspl 2501 Hmrl 364 Aell 2297 Bcetl 387 Apel - Ecoti 396 Banll - Sacl - EcosKI 402 lecs1 - Kpnl 408 Xmnl 2294 laczu Begl 2215 MCS wal - BoBI - Smal- Scal 2177 TspMI - Xmal 412 Hamill 417 thal 23 Prul 2066 Aecl - linell - Sall 429 Avall 2059 PUC19 2,686 bp Sh 434 Pstl 35 BaDI 1935 Acll 1924 Fipl 1919 Sphl 441 Hindl 417 Prull 628 wall 1837 NmeI 1822 Bell 1813 Bpml 1784 Bsrfl 1779 Bsal 1766 Til 641 BsaXI 659 Bspol - Sapl 683 Thl 781 BarD 1753 Bnrl 1744 ANIII - Peil 006 Drdl 908 ori Ahdl 1694 BeiVI 1015 Bsell 1110 AlwNI 1217 kell 1292 A. Which plasmid is used? B. Which antibiotic was used?

Human Anatomy & Physiology (11th Edition)
Human Anatomy & Physiology (11th Edition)
11th Edition
ISBN:9780134580999
Author:Elaine N. Marieb, Katja N. Hoehn
Publisher:Elaine N. Marieb, Katja N. Hoehn
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A. Read the following section to review what you learned about plasmids, transformation and antibiotic selection (which antibiotic?) Plasmids and Recombinant DNA Technology A plasmid is a circular piece of extra-chromosomal DNA (between 2000-10,000 bp) that is normally found in bacteria. In nature, the plasmid will often contain a gene that encodes a protein that allows the bacteria to be resistant to antibiotics, or a protein that allows the bacteria to cause an infection. Plasmids were discovered by researchers in the late sixties, and it was quickly realized that they could be used to amplify a gene of interest to create recombinant DNA. The gene of interest is inserted into the plasmid at restriction endonuclease sites. The plasmid can then be used as a vector to shuttle DNA to a different host/bacteria. There are many different kinds of plasmids commercially available. All of them contain: 1. a selectable marker (i.e., a gene that encodes for antibiotic resistance, color change, etc) 2. an origin of replication (which is used by the DNA making machinery in the bacteria as the starting point to make a copy of the plasmid) 3. a multiple cloning site (MCS). The multiple cloning site has many restriction enzyme sites and is used to insert the DNA of interest. The multiple cloning site is usually in the middle of a reporter gene like Lac Z. A commonly used plasmid that we will use in this lab is PUC19: (New England Biolabs) Indl 91 BamBI 51 BsmBl 2683 ISTAPI 179 Eco01091 2674 Ndel 183 Kasl - Narl - Sfol 235 Aatll - Zral 2617 Bell 245 Fspl 256 Prul 276 Pvull 306 BeVI 2542. Sspl 2501 Bmrl 364 Aell 2297 Boell 387 Apol - EcoRI 396 Hanll - Sacl - EcosSKI 402 Xmnl 2294 laczu lec651 - Kpnl 408 Begl 2215 MCS Awal - BaoRI - Smal - Scal 2177 TspMI - Xmal 412 Bamll 417 Nhal 423 Prul 2066 vall 2059 lel. Hindll. Sall 429 PUC19 2,686 bp Shf 434 BseI 1935 Pstl 435 Sphl 441 Adl 1924 Hindlll 417 Fspl 1919 Pvull 628 Avall 1837 NmeAlII 1822 Bell 1813 Til 641 BsaXI 659 Bpml 1784 Bsrll 1779 Bsal 1766 Bspol - Sapl 683 Til 781 AfAIII - Pcil 806 BsrDI 1753 Drdl 908 Bmrl 1744 ori Ahdl 1694 BeiVI 1015 BseYl 1110 AlwNI 1217 BeAl 1292 A. Which plasmid is used? B. Which antibiotic was used?
Transcribed Image Text:A. Read the following section to review what you learned about plasmids, transformation and antibiotic selection (which antibiotic?) Plasmids and Recombinant DNA Technology A plasmid is a circular piece of extra-chromosomal DNA (between 2000-10,000 bp) that is normally found in bacteria. In nature, the plasmid will often contain a gene that encodes a protein that allows the bacteria to be resistant to antibiotics, or a protein that allows the bacteria to cause an infection. Plasmids were discovered by researchers in the late sixties, and it was quickly realized that they could be used to amplify a gene of interest to create recombinant DNA. The gene of interest is inserted into the plasmid at restriction endonuclease sites. The plasmid can then be used as a vector to shuttle DNA to a different host/bacteria. There are many different kinds of plasmids commercially available. All of them contain: 1. a selectable marker (i.e., a gene that encodes for antibiotic resistance, color change, etc) 2. an origin of replication (which is used by the DNA making machinery in the bacteria as the starting point to make a copy of the plasmid) 3. a multiple cloning site (MCS). The multiple cloning site has many restriction enzyme sites and is used to insert the DNA of interest. The multiple cloning site is usually in the middle of a reporter gene like Lac Z. A commonly used plasmid that we will use in this lab is PUC19: (New England Biolabs) Indl 91 BamBI 51 BsmBl 2683 ISTAPI 179 Eco01091 2674 Ndel 183 Kasl - Narl - Sfol 235 Aatll - Zral 2617 Bell 245 Fspl 256 Prul 276 Pvull 306 BeVI 2542. Sspl 2501 Bmrl 364 Aell 2297 Boell 387 Apol - EcoRI 396 Hanll - Sacl - EcosSKI 402 Xmnl 2294 laczu lec651 - Kpnl 408 Begl 2215 MCS Awal - BaoRI - Smal - Scal 2177 TspMI - Xmal 412 Bamll 417 Nhal 423 Prul 2066 vall 2059 lel. Hindll. Sall 429 PUC19 2,686 bp Shf 434 BseI 1935 Pstl 435 Sphl 441 Adl 1924 Hindlll 417 Fspl 1919 Pvull 628 Avall 1837 NmeAlII 1822 Bell 1813 Til 641 BsaXI 659 Bpml 1784 Bsrll 1779 Bsal 1766 Bspol - Sapl 683 Til 781 AfAIII - Pcil 806 BsrDI 1753 Drdl 908 Bmrl 1744 ori Ahdl 1694 BeiVI 1015 BseYl 1110 AlwNI 1217 BeAl 1292 A. Which plasmid is used? B. Which antibiotic was used?
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