This page was created for an undergraduate assignment at Davidson College
Fragement Sizes That Will be Used to Determine the Orientation of the Insert
IDH1
length: 1083 nucleotides
EcoRV make the following cuts:
1. cuts plasmid 16bp from the insertion site
2. cuts IDH gene after nucleotide 340
Correct Orientation:
Fragment length: 356bp
|
Plasmid |
start------- IDH1 Gene (1083bp)-----------stop |
Plasmid |
| 16bp | 340bp | 743bp |
Reverse Orientation:
Fragment length: 759bp
| Plasmid | stop------- eneG 1 HDI(1083bp)-----------start | Plasmid |
| 16bp | 743bp | 340bp |
IDH2
length: 1110 nucleotides
BglII makes the following cuts:
1. cuts plasmid 15bp after the insertion site
2. cuts IDH gene after nucleotide 335
Correct orientation:
Fragment size: 790bp
| Plasmid | start------- IDH2 Gene (1110bp)-----------stop | Plasmid |
| 335bp | 775bp | 15 bp |
Reverse Orientation:
Fragmemt size 350bp
| Plasmid | stop------- eneG 2HDI(1110bp)-----------start | Plasmid |
| 775bp | 335bp | 15 bp |
IDP1
length: 1287 nucleotides
BglII makes the following cuts:
1. cuts plasmid 15bp after the insertion site
2. cuts IDP1 gene after nucleotide 1208
Correct orientation:
Fragment size: 94bp
| Plasmid | start------- IDP1 Gene (1287bp)-----------stop | Plasmid |
| 1208bp | 79bp | 15 bp |
Reverse Orientation:
Fragmemt size 1233bp
| Plasmid | stop------- eneG 2PDI(1287bp)-----------start | Plasmid |
| 79bp | 1208bp | 15 bp |
IDP2
length: 1239 nucleotides
BglII makes the following cuts:
1. cuts plasmid 15bp after the insertion site
2. cuts IDH gene after nucleotide 496
Correct orientation:
Fragment size: 758bp
| Plasmid | start------- IDP2 Gene (1239bp)-----------stop | Plasmid |
| 496bp | 743bp | 15 bp |
Reverse Orientation:
Fragmemt size 511bp
| Plasmid | stop------- eneG 2PDI(1239bp)-----------start | Plasmid |
| 743bp | 496bp | 15 bp |
IDP3
length: 1263 nucleotides
Two enzymes will be used:
1. HindIII cuts the plasmid 55 bp after the insertion site.
2. ClaI cuts the IDH gene after nucleotide 495.
Correct orientation:
Fragment size: 823 bp
| Plasmid | start------- IDP3 Gene (1263bp)-----------stop | Plasmid |
| 495 bp | 768bp | 55 bp |
Reverse Orientation:
Fragmemt size 550 bp
| Plasmid | stop------- eneG 3PDI(1263bp)-----------start | Plasmid |
| 768bp | 495bp | 55bp |