6. Double digest of all ligations: from previous days. Do a double digest to cut the components linearly, which will then be able to run through a gel to check if correct DNA is present. Follow the table below:
Parts | 10x Buffer | BSA | H20 | DNA | RE 1 | RE 2
|
L2a, LAMBDAcI:Term | 5.0uL | 0.5uL | 32.5uL | 10.0uL | EcoRI | Xba1
|
L2b, LAMBDAcI:Term | 5.0uL | 0.5uL | 32.5uL | 10.0uL | EcoRI | Xba1
|
L3i, Pro:LAMBDAcI | 5.0uL | 0.5uL | 32.5uL | 10.0uL | Pst1 | Spe1
|
L3j, Pro:LAMBDAcI | 5.0uL | 0.5uL | 32.5uL | 10.0uL | Pst1 | Spe1
|
L4b, LAMBDAcI:Term | 5.0uL | 0.5uL | 40.5uL | 2.0uL | EcoRI | Xba1
|
L4c, LAMBDAcI:Term | 5.0uL | 0.5uL | 39.5uL | 3.0uL | EcoRI | Xba1
|
L4d, LAMBDAcI:Term | 5.0uL | 0.5uL | 39.5uL | 3.0uL | EcoRI | Xba1
|
L4e, LAMBDAcI:Term | 5.0uL | 0.5uL | 39.5uL | 3.0uL | EcoRI | Xba1
|
L4g, LAMBDAcI:Term | 5.0uL | 0.5uL | 39.5uL | 3.0uL | EcoRI | Xba1
|
L4h, LAMBDAcI:Term | 5.0uL | 0.5uL | 39.5uL | 3.0uL | EcoRI | Xba1
|
L4i, LAMBDAcI:Term | 5.0uL | 0.5uL | 37.5uL | 5.0uL | EcoRI | Xba1
|
L4j, LAMBDAcI:Term | 5.0uL | 0.5uL | 32.5uL | 10.0uL | EcoRI | Xba1
|
L5c, GFP:Term | 5.0uL | 0.5uL | 40.5uL | 2.0uL | EcorI | Xba1
|
L6a, Pro:LAMBDAcI:Term | 5.0uL | 0.5uL | 39.5uL | 3.0uL | EcoRI | Xba1
|
L6b, Pro:LAMBDAcI:Term | 5.0uL | 0.5uL | 41.5uL | 1.0uL | EcoRI | Xba1
|
|