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(NOTE: Make sure that Buffer and Enzyme added last, enzyme after buffer)
| Add | Conc | Volm | Order of adding |
| DNA1 | c1 ng/ul | x1 = 50/c1 ul (50ng) | 2 |
| DNA2 | c2 ng/ul | x2 = 50/c2 ul (50ng) | 2 |
| GGDonr | c3 ng/ul | y = 50/c3 ul (100ng) | 2 |
| 10x T4 Ligase Buffer | 2ul | 3 | |
| BSA | 2ul | 3 | |
| BsaI (enzyme) | 1ul | 4 | |
| T4 Ligase (enzyme) | 1ul | 3 | |
| H20 | (20 - x1 - x2) ul | 1 | |
| Total | 20ul |
Take a p20, set it to 10uL and then pipet up and down.
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