Paul.xu at 12:39, 24 June 2014

2014-06-24T12:39:45Z

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== 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

{| width="200" cellspacing="1" cellpadding="1" border="1"
|-
| 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

[[Image:PEI800 DSP DTBP 6 21 2014.jpg]]

6/22/14

[[Image:PHP-DTBP-PEI800 6 22 2014.JPG]]

[[Image:PHP-DTBP-PEI800 6 22 2014 2.JPG]]

6/23/14

[[Image:PHP-DTBP-PEI18_6_23_2014.JPG]]

[[Image:PHP-DTBP-PEI18_6_23_2014_2.JPG]]

6/24/14

[[Image:PHP-DTBP-PEI18_6_24_2014.JPG]]

[[Image:PHP-DTBP-PEI18_6_24_2014_2.JPG]]

== Conclusion ==

Looks like DTBP is better.

[[Category:Transfection]] [[Category:Paul]]

Synthesis PHP-DTBP-PEI800 - Revision history

Paul.xu at 12:39, 24 June 2014