pkan-r/para sequence biotechnology lab program laboratory protocols by: marty ikkanda powerpoint by:...
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pKAN-R/pARA Sequence
Biotechnology Lab Program
Laboratory Protocols by:
Marty IkkandaPowerpoint by:
Anthony DauloPierce College, Woodland Hills, CA
V.1.2.4
Bruce Wallace
Restriction analysis of pKAN-R and pARA
pKAN-R
pARA
BamH I
Hind III
BamH IHind III
rfp702 bp
5408 bp
4058 bp
40 bp
Bruce Wallace
Restriction analysis of pKAN-R and pARA
BamH I Hind III
702bp
40 bp
BamH I Hind III
BamH IHind III
BamH IHind III
Restriction fragments after digest with Hind III and BamH I
4018 bp
4706 bp
Bruce Wallace
Ligation of pKAN-R/pARA restriction fragments
sticky endBamH I
sticky endHind III
sticky endBamH I
sticky endHind III
3’
5’
5’
3’
3’
3’
5’
5’5’
5’3’
3’
Bruce Wallace
Ligation of pKAN-R/pARA restriction fragments
Recombinant plasmid of interest
pARA-R
BamH I
Hind III
rfp702bp
4720 bp
Bruce Wallace
Preparing competent cells for transformation
Lipid bilayer(inner)
Lipid bilayer(outer)
Peptidoglycanlayer
Adhesion zone
Calcium ions
Bruce Wallace
Calcium ions
pARA-R
Transforming Escherichia coli with pARA-R
Adhesion zone
Lipid bilayer(inner)
Lipid bilayer(outer)
Peptidoglycanlayer
Bruce Wallace
Growth of transformed bacteria on various plates
P+ plates
P- plates
LB LB/amp LB/amp/ara
LB LB/amp
No growth
Bruce Wallace