Synthesis PHPCBA-DSP-PEI12

0.75 g 1,3-diamino-2-propanol (solid), 8.4 mmole

+ 1.5 ml MeOh to dissolve

 #1 + 50 mg CBA 

 #2 + 100 mg CBA 

+ 1.84 g 1, 3-dibromo-2-propanol liquid (~ 1ml), 8.4 mmole

react at 50 oc for 2 days.

+ 5 ml H2O to dissolve, +30 ml EtOH, + 10 ml isopropano, spin, remove solvent. wash again.

 to #1, + 60 mg dsp + 40 mg PEI12

to #2 + 40 mg dsp + 40 mg PEI12

react at 37 oC o/n.

resulting product:

#1: PHPCBA50-DSP-PE 

#2: PHPCBA100-DSP-PEI

transfection, 1 ug DNA, 3 ul diluted polymer

Plate 1

PHP-DSP-PEI18, 1:10	PHP-DSP-PEI18, 1:20	PHP-DSP-PEI18, 1:30	PHP-DSP-PEI18, 1:40
PHPCBA50-DSP-PEI, 1:10	PHPCBA50-DSP-PEI, 1:20	PHPCBA50-DSP-PEI, 1:30	PHPCBA50-DSP-PEI, 1:40
PHPCBA100-DSP-PEI, 1:10	PHPCBA100-DSP-PEI, 1:20	PHPCBA100-DSP-PEI, 1:30	PHPCBA100-DSP-PEI, 1:40

picture 16 hr after transfection (picture taken at 9AM on Nov, 28, 2011, transfection: 5PM on Nov 27, 2011)

11/29/2011

11/30/2011

2011/12/1

Plate 2

PHP-DSP-PEI18, 1:50	PHP-DSP-PEI18, 1:60	PHP-DSP-PEI18, 1:70	PHP-DSP-PEI18, 1:80
PHPCBA50-DSP-PEI, 1:50	PHPCBA50-DSP-PEI, 1:60	PHPCBA50-DSP-PEI, 1:70	PHPCBA50-DSP-PEI, 1:80
PHPCBA100-DSP-PEI, 1:50	PHPCBA100-DSP-PEI, 1:60	PHPCBA100-DSP-PEI, 1:70	PHPCBA100-DSP-PEI, 1:80

picture 16 hr after transfection (picture taken at 9AM on Nov, 28, 2011, transfection: 5PM on Nov 27, 2011)

11/29/2011

2011/11/30

2011/12/1

Plate 3

PHP-DSP-PEI18, 1:10	PHP-DSP-PEI18, 1:20	PHP-DSP-PEI18, 1:30	PHP-DSP-PEI18, 1:40
PHPCBA50-DSP-PEI, 1:10	PHPCBA50-DSP-PEI, 1:20	PHPCBA50-DSP-PEI, 1:30	PHPCBA50-DSP-PEI, 1:40
PHPCBA100-DSP-PEI, 1:10	PHPCBA100-DSP-PEI, 1:20	PHPCBA100-DSP-PEI, 1:30	PHPCBA100-DSP-PEI, 1:40

picture 16 hr after transfection (picture taken at 9AM on Nov, 28, 2011, transfection: 5PM on Nov 27, 2011)

11/29/11

2011/11/30

2011/12/1

Conclusion: Add CBA to polymer synthesis reaction will decrease transfection efficiency.