Synthesis PHP-DTBP-PEI800

Synthesis PEI800-DTBP and PEI800-DSP

6/18/2014

1. Make 400 mg/ml PEI800 in DMSO: 800 mg in 2 ml DMSO

2. Add 330 ul PEI800 in DMSO to 500 ul of 100 mg/ml DTBP in DMSO. React at 18 oC o/n, shaking. Form soluble dark brown liquid. Termed PEI800-DTBP: 0.48g/ml PEI00

3. Add 0.5 ml PEI800 in DMSO to 500 ul of 100 mg/ml DTBP in DMSO. React at 18 oC o/n, shaking. For some aggregate: Termed PEI800-DSP: 0.4 g/ml PEI800

6/19/2014

Use PHP suspension (6/15/14, 0.75g DAP+ 1.8g DBP) in 12 ml DMSO as backborne

Make DSP and DTBP 100 mg/ml in DMSO. DTBP is suspension

add volume: ul

Sample #	PHP suspension	PEI800-DTBP	PEI800-DSP	PEI1800	DSP 100mg/ml	DTBP 100mg/ml	Total
1	50	7.5			5
2	50	7.5				5
3	50		7.5		5
4	50		7.5			5
5	50			5	5
6	50			5		5
7	50	5			5
8	50	5				5
9	50		5		5
10	50		5			5

Need to add at least 5 ul of PEI800-DTBP to 50 ul PHP suspension to make the mixture clear.

Transfection

6/20/2014

Dilute in 10 mM Hepes-NaOH, pH 7.0

use 1 ug DNA, 3 ul diluted TR, 25 ul DMEM, 24 well plate, 293T/293E mix

6/22/14

6/23/14

6/24/14

Conclusion

Looks like DTBP is better.