Sites of restriction enzyme digestion are indicated by a slash AatI AGG/CCT BamH
ID: 33357 • Letter: S
Question
Sites of restriction enzyme digestion are indicated by a slash
AatI AGG/CCT BamHI G/GATCC BbrPI CAC/GTG
ClaI AT/GCAT EcoRI G/AATTC HinPI G/CGC
MboI /GATC MunI C/AATTG PmeI GTTT/AAAC
PstI CTGCA/G SacI GAGCT/C Sau3AI /GATC
SmaI CCC/GGG SnaBI TAC/GTA TaqI T/CGA
XbaI T/CTAGA XmnI GAANN/NNTTC
1) Use the table above to find two pairs of restriction enzymes that leave ends compatible for ligation with each other. (i.e. ends made by RE 1 can be ligated to ends made by RE 2).
2) You want to ligate a DNA fragment containing the insulin gene to a plasmid so you can make insulin in bacterial cells. The plasmid (not shown) has unique restriction sites for AatI, BamHI and MunI in its polylinker. The map below shows restriction sites around the insulin gene (the box). How would you digest each DNA for the ligation?
Explanation / Answer
Sac I and pst I can be used as RE1 and RE11 for ligation. these two will be compatible for after cutting ligation.
2. as the plasmid has mun I and bamH I site in its MSC, digest the plasmid DNA with Mun I and Bam H I