IGEM:Harvard/2006/DNA nanostructures/Notebook/2006-8-17

2nd PEG Precipitation

• conduct 2nd PEG precipitation on yesterday's precipitated samples (5.0A, C, D 7%-10%)
• protocol: mix all ingredients, add nanostructures last, total volume is 100 mL, incubate on ice for 15 min, spin for 10 min at 16k rcf, pipet off supernatant, resuspend pellet in 100 μL water

Final PEG %	PEG μL	NaCl μL	c5.0 μL	dH2O μL
7	35	10	40	15
8	40	10	40	10
9	45	10	40	5
10	50	10	40	0

lane	contents	loading dye
1	8 μL untreated 5.0.A	3 μL
2	20 μL 5.0.A in 7% PEG and 0.5 M NaCl (supernatant)	3 μL
3	8 μL 5.0.A in 7% PEG and 0.5 M NaCl (pellet)	3 μL
4	20 μL 5.0.A in 8% PEG and 0.5 M NaCl (supernatant)	3 μL
5	8 μL 5.0.A in 8% PEG and 0.5 M NaCl (pellet)	3 μL
6	20 μL 5.0.A in 9% PEG and 0.5 M NaCl (supernatant)	3 μL
7	8 μL 5.0.A in 9% PEG and 0.5 M NaCl (pellet)	3 μL
8	20 μL 5.0.A in 10% PEG and 0.5 M NaCl (supernatant)	3 μL
9	8 μL 5.0.A in 10% PEG and 0.5 M NaCl (pellet)	3 μL
10	1 kb+ ladder	3 μL
11	2.7 μL p7308	3 μL
12	8 μL untreated 5.0.C	3 μL
13	20 μL 5.0.C in 7% PEG and 0.5 M NaCl (supernatant)	3 μL
14	8 μL 5.0.C in 7% PEG and 0.5 M NaCl (pellet)	3 μL
15	20 μL 5.0.C in 8% PEG and 0.5 M NaCl (supernatant)	3 μL
16	8 μL 5.0.C in 8% PEG and 0.5 M NaCl (pellet)	3 μL
17	20 μL 5.0.C in 9% PEG and 0.5 M NaCl (supernatant)	3 μL
18	8 μL 5.0.C in 9% PEG and 0.5 M NaCl (pellet)	3 μL
19	20 μL 5.0.C in 10% PEG and 0.5 M NaCl (supernatant)	3 μL
20	8 μL 5.0.C in 10% PEG and 0.5 M NaCl (pellet)	3 μL
21	8 μL untreated 5.0.D	3 μL
22	20 μL 5.0.D in 7% PEG and 0.5 M NaCl (supernatant)	3 μL
23	8 μL 5.0.D in 7% PEG and 0.5 M NaCl (pellet)	3 μL
24	20 μL 5.0.D in 8% PEG and 0.5 M NaCl (supernatant)	3 μL
25	8 μL 5.0.D in 8% PEG and 0.5 M NaCl (pellet)	3 μL
26	20 μL 5.0.D in 9% PEG and 0.5 M NaCl (supernatant)	3 μL
27	8 μL 5.0.D in 9% PEG and 0.5 M NaCl (pellet)	3 μL
28	20 μL 5.0.D in 10% PEG and 0.5 M NaCl (supernatant)	3 μL
29	8 μL 5.0.D in 10% PEG and 0.5 M NaCl (pellet)	3 μL
30	1 kb+ ladder	3 μL
31	2.7 μL p7308	3 μL
32-40	(empty)	3 μL