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interactome:experiments:20191220 [2019/12/20 09:42] dan |
interactome:experiments:20191220 [2019/12/20 09:45] (current) dan |
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| *MORF4L1-40bp overlap for all genes | *MORF4L1-40bp overlap for all genes | ||
| *5 pilot genes with 2 barcodes each. | *5 pilot genes with 2 barcodes each. | ||
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| + | *Frd primer : BCamplif-Frd-MORF4L1-v2-4;PI-34-BClib-reamp-MRFAP1-Frd;PI-35-BClib-reamp-BCL2L2-Frd;PI-36-BClib-reamp-BIK-Frd;PI-37-BClib-reamp-MRFAP1L1-Frd | ||
| + | *Rvs primer : PI-33-Barcode-reamp1-Rvs | ||
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| | | Fusion PCR template | 0.5 | 1 | | | | | | | Fusion PCR template | 0.5 | 1 | | | | | ||
| | ^ Total (µL) ^ 10 ^ 20 | | | | | | ^ Total (µL) ^ 10 ^ 20 | | | | | ||
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| ^ STEP ^ Action ^ Time ^ Memo ^ | ^ STEP ^ Action ^ Time ^ Memo ^ | ||
| | 1 | 98˚C | 3 min | Initial denaturation. | | | 1 | 98˚C | 3 min | Initial denaturation. | | ||
| - | | 2 | 98˚C | 10 sec | Denaturation. | | + | | 2 | 98˚C | 30 sec | Denaturation. | |
| | 3 | 65 ˚C | 30 sec | Annealing. See Tm table | | | 3 | 65 ˚C | 30 sec | Annealing. See Tm table | | ||
| - | | 4 | 72˚C | 3 min | Extension. **15sec/kbp, but using longer** | | + | | 4 | 72˚C | 30 sec | Extension. **15sec/kbp, but using longer** | |
| | 5 | GOTO 2 | x 29 | | | | 5 | GOTO 2 | x 29 | | | ||
| | 6 | 72˚C | 5:00 | Final extension. | | | 6 | 72˚C | 5:00 | Final extension. | | ||