Timeline (week #): 2 E coli transformation - must return one day later for analy

Question

Timeline (week #): 2 E coli transformation - must return one day later for analysis 3 Plasmid isolation, quantification, and restriction enzyme digestion Note: we will also begin trial B-galactosidase assays this week 5 Postlab due 4 Complete plasmid analysis using gel electrophoresis Specifie background information for our experiments Plasmids are circular pieces of DNA that are extra-chromosomal (not part of the chromosome, but independently replicated and transferred to progeny). Plasmids can be present in many (even hundreds) of copies in a single cell, making them very useful for DNA studies. You will receive one of two plasmids; the identity unknown to you. It will most likely be pBR322 or pUC19. Below is a plasmid map for each that shows the major features of the plasmid, including the genes and promoter regions, as well as recognition sites for several restriction enzymes. Bsu151 23 EcoRI, Xapl 4350, |Hindill 29 Eco321 185 The orange text represents the locations of unique restriction endonuclease sites. The numbers indicate the relative locations of the restriction Aatil 4284 Nhel 220 BamlI 3TS Tsti 289 Scal 3344 502 Pvul 153 Sall 651 Pst Vspl Boxl 712 Eco311 343 Eam11051 3361 Eco521 939sites. This plasmid Bsp681 972 has 4361 bp, and Bvel 1083 PvulI cuts it at bp pBR322 4361 bp 2064. Mva12691 1353 Eco1301 1309 Eco88l 1425 Misl 1444 Call 2884 Bpu1ol 1580 Bopl 1650 Al, Pscl 2473 Kpn21 1004 BseJl 1068 Pvull 2004 2117 Ndel 2295 Bst11071 2244 Esp3l 2122 Ppu21l 2225l IPsyl 2217

Explanation / Answer

D4.

Size fragments generated after digestion with Scal and HindIII

Plasmid pBR322 is 4361bp, it has Scal restriction site at 2844 bp and Hindlll has at 29bp So after digestion with both enzyme there is cut at 29, 2844 bp

So it produce 2 fragment of size 1546bp and 2815

Plasmid pUC19 is 2686bp long plasmid , Scal has restriction site at 1191bp and Hindlll at 233bp

So it produce 958 bp and 1728bp fragment

And these two plasmids can easily identify after digestion due to there different fragment size.

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