IGEM:Harvard/2006/vlau/Notebook/2006-7-25
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Pre-Working Stocks & Working Stocks
1. Rxn Mixtures
Description | Plate Locations | Total |
c4.0.2: core top lid oligos | box 4_A: row F (7-12), row G (1-12), row H (1-12); box 4_B: row A (1-3) | 33 |
c4.0.3: core bottom lid oligos | box 4_B: row A (4-12), row B (1-12), row C (1-12) | 33 |
Working Stocks | Description | Pre-Working Stocks | Water | Total |
c4.0Fb.core | latch design 1, inside biotin | 1 (66 μL, 2 (33 μL), 3 (33 μL), 4b (4 μL) | 64 μL | 200 μL |
c4.0Fb.latches | latch design 1, inside biotin | 8 (4 μL), 11 (2 μL), 12 (2 μL) | - | - |
c4.0Gb.core | latch design 1, outside biotin | 1 (66 μL, 2 (33 μL), 3 (33 μL), 6b (4 μL) | 64 μL | 200 μL |
c4.0Gb.latches | latch design 1, outside biotin | 8 (4 μL), 11 (2 μL), 12 (2 μL) | - | - |
c4.0Hb.core | latch design 2, inside biotin | 1 (66 μL, 2 (33 μL), 3 (33 μL), 4b (4 μL) | 64 μL | 200 μL |
c4.0Hb.latches | latch design 2, inside biotin | 9 (4 μL), 13 (2 μL), 14 (2 μL), 23 (4 μL), 24 (2 μL), 25 (2 μL) | - | - |
c4.0Ib.core | latch design 2, outside biotin | 1 (66 μL, 2 (33 μL), 3 (33 μL), 6b (4 μL) | 64 μL | 200 μL |
c4.0Ib.latches | latch design 2, oustide biotin | 9 (4 μL), 13 (2 μL), 14 (2 μL), 23 (4 μL), 24 (2 μL), 25 (2 μL) | - | - |
2. Protocol
- mixed 10 μL of each 50 μM stock oligo to make pre-working stocks - mixed listed pre-working stocks to make working stocks where final concentration of each oligo is 250 nM
Folding Rxns w/ Biotinylated Oligos
1. Reagents (for c4.0Fb, c4.0Gb, c4.0Hb, c4.0Ib)
- 9 μL p7308 scaffold = (10 nM)/(44 nM) * 40 μL - 16 μL oligos = (100 nM)/(250 nM) * 40 μL - 4 μL 10x folding buffer (500 mM HEPES pH 7.5, 500 mM NaCl, 100 mM MgCl2) - 11 μL dH2O - total volume: 40 μL
2. Negative Controls
- 1st (no oligos): 9 μL scaffold, 16 μL dH2O, 4 μL buffer, 11 μL dH2O - 2nd (no scaffold): 9 μL dH2O, 16 μL oligos, 4 μL buffer, 11 μL dH2O