4 Name: Date: Period: E Plasmid Mapping Practice Worksheet #1 AP Biology 1. Below is a restriction map for the plasmid pGEN101 (total length = 20 kb). Using this map as a guide, give the number of restriction fragments along with their associated lengths that would result from digesting pGENIO1 with the restriction enzymes EcoRI, BamHI, and a combination of EcoRI + BamHI. kb BamHI BamHI 2kb, BamHI. kb Ae EcoRI Digest Performes EeoRL.. BamHl. EcoRI + BamHl......-.see0+ 20 Ke 2ikb, (obo, IL Kb Sizes of Fragments Obtained: Zico, Hk, @kb, Oko ‘Two freshmen college students, interested in becoming gene jocks, performed the following set of restriction digests on a newly isolated plasmid, pBLA230. The reaction they carried out, along with the fragment obtained in single and double digest reactions were: Enzyme(s) Fragment Lengths Obtained: Hpal 26 kb Hindi 13 kb, 6 kb, 4 kb, 3 kb Hpal + Hindi Tb, 6 kb (2), 4kb, 3kb Using this information, construct a restriction map of pBLA230. Wid TIL : Hid nName’ Date Period: AP Biology pBR607 is a 2.6 kb plasmid containing Amp in and Tetracycline resistance markers, an origin of replication, and unique restriction sites for the restrietion enzymes EcoRI, BamHI, and PstI. Given the restriction map for pBR607 for the enzymes EcoRI, BamHI, and PstI, show on the agarose gel picture below where the approximate positions of the restriction fragments, generated from the given restriction digests would be located after carrying out electrophoresis. BamHI 0.2 kb EcoRI 1.94kb 144 eee (0.46 kb Zu0 Pstl Size FeoRI& EcoRI & Standards Pst Bam 4.0kb | ——— “ae 2.2kb 20kb | ——— 05k | ——— __ rr ie All add up fo Kwon lergts (70k)Name: Date: Period: AP Biology As part of an undergraduate project, a student was attempting to construet a restriction map for the plasmid pUC23 using the restriction enzymes EcoRI and BamAHl. After carrying out both single and double enzyme digest reactions and electrophoresing each reaction mix through an aragose gel, the picture below is obtained, showing the number of DNA fragments produced in each reaction, along with the sizes of each fragment, From this information, construct a restriction map of the pUC23 for enzymes EcoRI and BamHl. EcoRI & he aack EcoRI Bamiil BamHI 2 bands bee ain HE ?Name: Date: Period: 5. Avery determined graduate student set out (0 construct a restriction map for the plasmid pDA4O1 (total size = 4.0 kb). The restriction enzymes used were HindIII, BamHI, and EcoRI. After carrying out the digest AP Biology ns, the resulting DNA fragments were electrophoresed and sized using a set of DNA size standards. The data obtained in cach digestion are shown below. From this data, construct a restriction map of pDAdO1 for the enzymes HindIII, BamHI, and EcoRI. Enzyme(s): 43 HindiIl== 3 | BamHI 4 yt” &| PeoRT = x” HindIII + EcoRI iw Ts Bamlll + EcoRT dP boat 3 eke aS ko yaske 3 ase Segments observed after digestion: 3.82 kb, 0.18 kb 35 kb, 1.65 kb 3.00 kb, 1.00 kb jae Hind/L+ BamHI 2.35 kb, 1.20 kb, 0.27 kb, 0.18 kb 1.87 kb, 1.00 kb, 0.95 kb, 0.18 kb: 1.60 kb, 1.40 kb, 0.75 kb, 0.25 kh as at at el BS » 2.35 Bt yee‘Name: Date: Period AP Biology 6. Acircular DNA plasmid, pDA102, has a size of 4.35 kb. When the plasmid DNA digested with combinations of restriction enzymes and the resulting fragments are electrophoresed, the following data is obtained. Using these data, construct a restriction map of plasmid pDA102 for the restriction enzymes Sal and Hhalll, Restriction Enzyme(s)<> Sall ‘haut Sall + Hhallt 45 OF +o te make pustakes!