2
edits
LIC cloning: Difference between revisions
Jump to navigation
Jump to search
no edit summary
(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...) |
No edit summary |
||
| (One intermediate revision by the same user not shown) | |||
| Line 11: | Line 11: | ||
{| border="1" cellpadding="5" cellspacing="0" | {| border="1" cellpadding="5" cellspacing="0" | ||
|- | |- | ||
| | |forward | ||
|cagggacccggtAGTGCTGCAGTGACTGC | |cagggacccggtAGTGCTGCAGTGACTGC | ||
|- | |- | ||
| | |reverse | ||
|cgaggagaagcccggttaTGGTTCTTCTTTTTCCCGGGG | |cgaggagaagcccggttaTGGTTCTTCTTTTTCCCGGGG | ||
|- | |- | ||
| Line 150: | Line 150: | ||
<br> | <br> | ||
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> | ||