| Date | Product | Gene | Starting Plasmid | Forward PrimerReverse Primer | Thermocycler Variables: #cycles / annealing temp / extension time | Gel Picture | |||
|---|---|---|---|---|---|---|---|---|---|
| 6/23 | Q2-BACE2-QX | BACE2 | pEXPR BACE2 | iGEM 016 BACE2 C-term primerF | iGEM 017 BACE2 C-term primerR | 30 / 60C / 30s | Gel Picture 6/23 | ||
| Q1-eYFP-Q2eYFP | pEXPR TRE:eYFPiGEM 018 eYFP N-term primerF | iGEM 019 eYFP N-term primerR | |||||||
| Q1-BACE2-Q2BACE2 | pEXPR BACE2 | iGEM 001-2 BACE2 N-term primerF | iGEM 002-2 BACE2 N-term primerR | ||||||
| Q2-eYFP-QXeYFP | pEXPR TRE:eYFPiGEM 003-2 YFP C-term primerF | iGEM 004-2 YFP C-term primerR | |||||||
| 6/24 | Q2-BACE2-QX | BACE2 | pEXPR BACE2 | iGEM 016 BACE2 C-term primerF | iGEM 017 BACE2 C-term primerR | 35 / 56.2C / 30s | Gel Picture 6/24 | ||
| Q1-eYFP-Q2eYFP | pEXPR TRE:eYFP | iGEM 018 eYFP N-term primerF | iGEM 019 eYFP N-term primerR | ||||||
| Q1-BACE2-Q2BACE2 | pEXPR BACE2 | iGEM 001-2 BACE2 N-term primerF | iGEM 002-2 BACE2 N-term primerR | ||||||
| Q2-eYFP-QXeYFP | pEXPR TRE:eYFP | iGEM 003-2 YFP C-term primerF | iGEM 004-2 YFP C-term primerR | ||||||
| 6/25 | Q1-BACE2-Q2 | BACE2 | pEXPR BACE2 | iGEM 001-2 BACE2 N-term primerF | iGEM 002-2 BACE2 N-term primerR | 35 / 50.2C / 30s
| Gel Picture 6/25 | ||
| Q2-BACE2-QXBACE2 | pEXPR BACE2iGEM 016 BACE2 C-term primerF | iGEM 017 BACE2 C-term primerR | |||||||
| Q2-BACE2-QX | BACE2 | pEXPR BACE2 | iGEM 016 BACE2 C-term primerF | iGEM 017 BACE2 C-term primerR | |||||
| Q1-eYFP-Q2eYFP | pEXPR TRE:eYFPiGEM 018 eYFP N-term primerF | iGEM 019 eYFP N-term primerR | |||||||
| Q2-eYFP-QX | eYFP | pEXPR TRE:eYFP | iGEM 003-2 YFP C-term primerF | iGEM 004-2 YFP C-term primerR | |||||
| 6/26 | Q2-BACE2-QX BACE2(plus betaine) | pEXPR BACE2 | iGEM 016 BACE2 C-term primerF | iGEM 017 BACE2 C-term primerR | 30 / 55C / 1m
| Gel Picture 6/26 | |||
Q2-BACE2-QX BACE2(plus DMSO) | pEXPR BACE2iGEM 016 | ||||||||
Q1-BACE2-Q2 (plus betaine) | pEXPR BACE2 C-term primerFiGEM 017 BACE2 C-term primerR | ||||||||
Q1-BACE2-Q2 BACE2(plus DMSO) | pEXPR BACE2 | iGEM 001-2 BACE2 N-term primerF | iGEM 002-2 BACE2 N-term primerR | ||||||
| Q1-BACE2-Q2 | BACE2 | pEXPR BACE2 | iGEM 001-2 BACE2 N-term primerF | iGEM 002-2 BACE2 N-term primerR | |||||
| 6/27 | Q2-BACE1-QX | pEXPR BACE1 | Gel Picture 6/27 | ||||||
| Q2-eYFP-QX | pEXPR eYFP | ||||||||
| 6/30 | Q1-eYFP-Q2 | pEXPR eYFP | Gel Picture 6/30 | ||||||
| Q1-BACE1-Q2BACE2 | pEXPR BACE1 | iGEM 014 BACE1 N-term primerF | iGEM 015 BACE1 N-term primerR | ||||||
| Q2-BACE1-QX | BACE2 | pEXPR BACE1 | iGEM 023 BACE1 C-term primerF | iGEM 022 BACE1 C-term primerR | |||||
| Q1-eYFP-Q2 | eYFP | pEXPR eYFP | iGEM 021 eYFP N-term primerF | iGEM 020 eYFP N-term primerR | |||||
| 7/9 | Q2-eYFP-QX (for BACE2) | pEXPR eYFP | 35 / 58C / 1m with both DMSO and Betaine | Gel Picture 7/9 | |||||
| Q1-eYFP-Q2 (for BACE2) | pEXPR eYFP | ||||||||
| Q2-eYFP-QX (for BACE1) | pEXPR eYFP | ||||||||
| Q2-BACE2-QX | pEXPR BACE2 | ||||||||
| Q1-BACE2-QX | pEXPR BACE2 | ||||||||
| Q1-BACE1-QX | pEXPR BACE1 | ||||||||
| 7/10 | (miRNA generator stuff) | 35 / 58C / 1m with both DMSO and Betaine | Gel Picture 7/10 | ||||||
| (20 PCR products in all) | |||||||||
| 7/11 | (miRNA generator stuff) | 30 / 61C / 30s | Gel Picture 7/11 | ||||||
| (12 PCR products in all) | |||||||||
| 7/16 | (miRNA generator stuff) | 30 / 63C / 30s | |||||||
| (12 PCR products in all) | Q2-eYFP-QX | eYFP | pEXPR eYFP | iGEM 012 eYFP C-term primerF | iGEM 013 eYFP C-term primerR |
Ran PCRs according to protocol on NEB website: https://www.neb.com/protocols/2012/09/06/protocol-phusion-high-fidelity-pcr-master-mix-with-hf-buffer-m0531
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