1. DNA can be cut at specific sequence motifs using enzymes called restriction endonucleases. You obtain a single PCR product from R. sphaeroides DNA that can be cut in the places shown below by solid arrows. These are the Clal specific restriction motifs. Positions of these restriction sites are indicated by nucleotide numbers in a 1059 nucleotide DNA fragment. Draw the expected band(s) for each of the bacterial species on the gel picture below if treated with the restriction enzymes as illustrated. Rhodobacter sphaeroides 2. 4.1 1059 0 290 749 Rhodopseudomonas palustris BisB5 0 Dinoroseobacter shibae DFL 12 0 290 Silicibacter sp. TM 1040 0 1059 749 1059 1059 DNA ladder R. sphaeroides 2. 4. 1 D. shibae DFL12 R. palustris BisB5 Silicibacter sp. TM1040 1100 1000 900 800 700 600 500 400 300 200 100 |||||||| CS Scanned with CamScanner 54

DNA can be cut at specific sequence motifs using enzymes called restriction endonucleases. You obtain a single PCR product from R. sphaeroides DNA that can be cut in the places shown below by solid arrows. These are the Clal specific restriction motifs. Positions of these restriction sites are indicated by nucleotide numbers in a 1023 nucleotide DNA fragment. Draw the expected band(s) for each of the bacterial species on the gel picture below if treated with the restriction enzymes as illustrated. Rhodobacter sphaeroides 2. 4. 1 0. 1023 150 627 Rhodopseudomonas palustris BisB5 0. 1023 627 Dinoroseobacter shibae DFL 12 1023 150 Silicibacter sp. TM 1040 1023 DNA ladder R sphaeroides 2. 4. 1 R palustris BisB5 D. shibae DFL12 Silicibacter sp. TM1040 1100 1000 900 800 700 600 500 400 300 200 100

The map of plasmid pUC19 is shown below.  The restriction site coordinate is the position of the 5’base on the top strand of each site sequence.  The restriction enzyme sites are in bold type if there is only one site in pUC19. Please list the fragments in order of size, largest to smallest, which will result from a complete digestion by the restriction enzyme PvuII. Please list the fragments in order of size, largest to smallest, which will result from a complete digestion by the restriction enzyme DrdI.

Kha Vu Danels Include: 8DX : Safehy Jor bromie tnto lab repart! Name Section date sheet MAPPING PRACTICE #1 Below is a restriction map for the plasmid PGEN 101 (total length = 20 Kb). Using this map as a guide, give the number of restriction fraqments along with their associated lengths that would result from digesting PGEN 101 with the restriction enzymes EcoRI, BamHI and a combination of ECORI and BamHI. BamHI 3.2 Kb 1.7 Kb EcoRI BamHI PGEN 101 8.7 Kb 5.5 Kb .9 Kb EcoRI ECORI DIGESTION PERFORMED SIZES OF FRAGMENTS OBTAINED 10.4 kb , 0.9kb, 8.7 Kb EcoRI 3.2 Kb, 16. 8kb BamHI EcoRI + BamHI