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LIC cloning: Difference between revisions
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(New page: '''LIC-cloning into the pET-NKI LIC vectors''' [contributed by Vangelis Christodoulou, NKI] This protocol is based on the "version 2" LIC system designed by Vangelis. '''DAY -20 to -10...) |
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Conjoin the vector and insert <br> | Conjoin the vector and insert <br> | ||
1) Add | 1) Add 2 uL of the T4 treated insert to a 1.5 mL eppendorf tube<br> | ||
2) Add | 2) Add 1 uL of the T4 treated vector (~50ng/uL)<br> | ||
3) Incubate together at 22oC for 5 minutes.<br> | 3) Incubate together at 22oC for 5 minutes.<br> | ||
4) Stop the reaction by adding 1 uL of sterile 25 mM EDTA.<br> | 4) Stop the reaction by adding 1 uL of sterile 25 mM EDTA.<br> | ||