Editing Gu PHP-DSP-PEI18 (section)

== Stability  ==

Dilute polymer PHP-Gu-PEI18 with DMSO or 10 mM HEPES, pH7.0&nbsp; to make&nbsp; 1:10 stock. 

Heat at 50 oC for 2 days. 

Use 1:10&nbsp; stock or 3-fold dilution&nbsp; (diluted 3 folds) for transfection 

{| cellspacing="1" cellpadding="1" border="1" width="514"
|-
| <br> 
| Sample 
| Transfection Dilution 
| Heat treatment 
| Storage solvent
|-
| colspan="5" | No heat<br>
|-
| 1 
| 6/24/14#1<br> 
| 1:30<br> 
| n 
| H2O
|-
| 7 
| 6/24/14#1 <br> 
| 1:30 
| n 
| DMSO
|-
| 13 
| 6/24/14#1 
| 1:10 
| n 
| H2O
|-
| 19 
| 6/24/14#1 <br> 
| 1:10&nbsp; <br> 
| n 
| DMSO
|-
| 2 
| 6/24/14#5P 
| 1:30 
| n 
| H2O <br>
|-
| 8 
| 6/24/14#5P 
| 1:30 
| n 
| DMSO<br>
|-
| 14 
| 6/24/14#5P<br> 
| 1:10 
| n 
| H2O <br>
|-
| 20 
| 6/24/14#5P<br> 
| 1:10 
| n<br> 
| DMSO
|-
| 3 
| 7/3/14#1<br> 
| 1:30 
| n<br> 
| H2O&nbsp; <br>
|-
| 9 
| 7/3/14#1<br> 
| 1:30 
| n<br> 
| DMSO
|-
| 15 
| 7/3/14#1<br> 
| 1:10 
| n<br> 
| H2O
|-
| 21 
| 7/3/14#1<br> 
| 1:10 
| n<br> 
| DMSO
|-
| 4 
| 7/3/14#2&nbsp;&nbsp; <br> 
| 1:30 
| n<br> 
| H2O
|-
| 10 
| 7/3/14#2 
| 1:30 
| n<br> 
| DMSO
|-
| 16 
| 7/3/14#2 
| 1:10 
| n<br> 
| H2O
|-
| 22 
| 7/3/14#2 
| 1:10 
| n<br> 
| DMSO
|-
| colspan="5" | heat at 50 oC for two days<br>
|-
| 5 
| 6/24/14#1<br> 
| 1:30 <br> 
| y<br> 
| H2O&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; <br>
|-
| 11 
| 6/24/14#1 
| 1:30 
| y<br> 
| DMSO&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; <br>
|-
| 17 
| 6/24/14#1 
| 1:10 
| y<br> 
| H2O
|-
| 23 
| 6/24/14#1 
| 1:10 
| y<br> 
| DMSO
|-
| 6 
| 6/24/14#5P<br> 
| 1:30 
| y<br> 
| H2O
|-
| 12 
| 6/24/14#5P 
| 1:30 
| y<br> 
| DMSO
|-
| 18 
| 6/24/14#5P 
| 1:10 
| y<br> 
| H2O
|-
| 24 
| 6/24/14#5P 
| 1:10 
| y<br> 
| DMSO
|-
| <br> 
| <br> 
| <br> 
| <br> 
| <br>
|-
| 25 
| 7/3/14#1<br> 
| 1:30 
| y<br> 
| H2O
|-
| 27 
| 7/3/14#1 
| 1:30 
| y<br> 
| DMSO
|-
| 29 
| 7/3/14#1 
| 1:10 
| y<br> 
| H2O
|-
| 31 
| 7/3/14#1 
| 1:10 
| y<br> 
| DMSO
|-
| 26 
| 7/3/14#2 <br> 
| 1:30 
| y<br> 
| H2O
|-
| 28 
| 7/3/14#2 
| 1:30 
| y<br> 
| DMSO
|-
| 30 
| 7/3/14#2 
| 1:10 
| y<br> 
| H2O
|-
| 32 
| 7/3/14#2&nbsp; 
| 1:10 
| y<br> 
| DMSO<br>
|-
| <br> 
| <br> 
| <br> 
| <br> 
| <br>
|-
| L3K 
| <br> 
| <br> 
| <br> 
| <br>
|-
| Turbofect 
| <br> 
| <br> 
| <br> 
| <br>
|}

<br> 

[[Image:PHP GU PEI stability 7 8 14.JPG]][[Image:PHP GU PEI stability 7 8 14 2.JPG]] 

7/9/14<br> 

[[Image:PHP GU PEI stability 7 9 14 2.JPG]][[Image:PHP GU PEI stability 7 9 14.JPG]] 

7/10/14 

[[Image:PHP GU PEI stability 7 10 14.JPG]][[Image:PHP GU PEI stability 7 10 14 2.JPG]] 

7/11/14<br> 

[[Image:PHP GU PEI stability 7 11 14.JPG]][[Image:PHP GU PEI stability 7 11 14 2.JPG]] 

7/12/14<br> 

[[Image:PHP GU PEI stability 7 12 14.JPG]][[Image:PHP GU PEI stability 7 12 14 2.JPG]]<br> 

7/13/14 

[[Image:PHP GU PEI stability 7 13 14.JPG]][[Image:PHP GU PEI stability 7 13 14 3.JPG]] 

7/14/14<br> 

=== [[Image:PHP GU PEI stability 7 14 14 2.JPG]][[Image:PHP GU PEI stability 7 14 14.JPG]]<br>  ===

=== Stablity Test 2<br>  ===

7/10/14<br> 

Dilute polymer PHP-Gu-PEI18 with DMSO or 10 mM HEPES, pH7.0 to make 1:10 stock. 

Heat at 50 oC for 4 days. 

Use 1:10 stock for transfection, 293T 24 wells, 1ug DNA, 3 ul TR<br> 

<br> 

{| cellspacing="1" cellpadding="1" border="1" width="628"
|-
| Sample #<br> 
| 6/24/14 sample #1<br> 
| 6/24/14 sample #5P (purified by ethano ppt)<br> 
| 7/3/14#1<br> 
| 7/3/14#2<br> 
| Storage Buffer: 10 mM HEPES, pH7<br> 
| Storage: DMSO<br>
|-
| 1<br> 
| *<br> 
| <br> 
| <br> 
| <br> 
| *<br> 
| <br>
|-
| 2<br> 
| *<br> 
| <br> 
| <br> 
| <br> 
| <br> 
| *<br>
|-
| 3<br> 
| <br> 
| *<br> 
| <br> 
| <br> 
| *<br> 
| <br>
|-
| 4<br> 
| <br> 
| *<br> 
| <br> 
| <br> 
| <br> 
| *<br>
|-
| 5<br> 
| <br> 
| <br> 
| *<br> 
| <br> 
| *<br> 
| <br>
|-
| 6<br> 
| <br> 
| <br> 
| *<br> 
| <br> 
| <br> 
| *<br>
|-
| 7<br> 
| <br> 
| <br> 
| <br> 
| *<br> 
| *<br> 
| <br>
|-
| 8<br> 
| <br> 
| <br> 
| <br> 
| *<br> 
| <br> 
| *<br>
|}

Picture of 7/12/14 

[[Image:PHP GU PEI stability 7 12 14 3.JPG]]<br> 

7/13/14 

[[Image:PHP GU PEI stability 7 13 14 2.JPG]] 

7/14/14 <br> 

[[Image:PHP GU PEI stability 7 14 14 3.JPG]]<br> 

7/16/14 <br> 

[[Image:PHP GU PEI stability 7 16 14 2.JPG]]<br> 

7/17/14 

[[Image:PHP GU PEI stability 7 17 14 2.JPG]] 

7/18/14 

[[Image:PHP GU PEI stability 7 18 14 2.JPG]] 

7/19/14 

[[Image:PHP GU PEI stability 7 19 14.JPG]] 

7/20/14 

[[Image:PHP GU PEI stability 7 20 14 1.JPG]]<br> 

=== Stability Test 3  ===

incubate polymers at 50 oC for 7 days, other conditions are as Stability Test 2<br> 

transfection is on 7/14/14<br> 

picture of 7/16/14 7/14/14<br> 

[[Image:PHP GU PEI stability 7 16 14.JPG]]<br> 

7/17/14 

[[Image:PHP GU PEI stability 7 17 14.JPG]] 

7/18/14 

[[Image:PHP GU PEI stability 7 18 14.JPG]] 

<br> 

7/19/14 

[[Image:PHP GU PEI stability 7 19 14 2.JPG]] 

7/20/14 

[[Image:PHP GU PEI stability 7 20 14.JPG]] 

<br> 7/10/14<br> 

<br> Dilute polymer PHP-Gu-PEI18 with DMSO or 10 mM HEPES, pH7.0 to make 1:10 stock. 

Heat at 50 oC for 4 days. 

Use 1:10 stock for transfection, 293T 24 wells, 1ug DNA, 3 ul TR<br> 

<br> 

=== Stability Test 5<br>  ===

8/1//14 <br> 

Stability Test Samples, has been stored at 50 oC for &gt; 2 weeks<br> 

{| cellspacing="1" cellpadding="1" border="1" width="628"
|-
| Sample #<br> 
| 6/24/14 sample #1<br> 
| 6/24/14 sample #5P (purified by ethano ppt)<br> 
| 7/3/14#1<br> 
| 7/3/14#2<br> 
| Storage Buffer: 10 mM HEPES, pH7<br> 
| Storage: DMSO<br>
|-
| 1<br> 
| *<br> 
| <br> 
| <br> 
| <br> 
| *<br> 
| <br>
|-
| 2<br> 
| *<br> 
| <br> 
| <br> 
| <br> 
| <br> 
| *<br>
|-
| 3<br> 
| <br> 
| *<br> 
| <br> 
| <br> 
| *<br> 
| <br>
|-
| 4<br> 
| <br> 
| *<br> 
| <br> 
| <br> 
| <br> 
| *<br>
|-
| 5<br> 
| <br> 
| <br> 
| *<br> 
| <br> 
| *<br> 
| <br>
|-
| 6<br> 
| <br> 
| <br> 
| *<br> 
| <br> 
| <br> 
| *<br>
|-
| 7<br> 
| <br> 
| <br> 
| <br> 
| *<br> 
| *<br> 
| <br>
|-
| 8<br> 
| <br> 
| <br> 
| <br> 
| *<br> 
| <br> 
| *<br>
|}

<br> 

<br> 

8/1//14 Transfection <br> 

1 ug DNA, 293T cells in 24 well plate<br> 

{| cellspacing="1" cellpadding="1" border="1" width="542"
|-
| Well<br> 
| Sample<br> 
| Description<br> 
| Note<br>
|-
| 1<br> 
| PHP-DSP-AG<br> 
| <br> 
| <br>
|-
| 2<br> 
| PHP-DTBP-AG<br> 
| <br> 
| <br>
|-
| 3<br> 
| PHP-DSP-AG, ppt<br> 
| <br> 
| <br>
|-
| 4<br> 
| PHP-DSP-AG, oil<br> 
| <br> 
| <br>
|-
| 5<br> 
| K-DBP-AG<br> 
| Lysine + DBP + AG<br> 
| very weak DNA binding <br>
|-
| 6<br> 
| K-CBA-AG<br> 
| Lysine + CBA + AG<br> 
| very weak DNA binding <br>
|-
| 7<br> 
| PHP-DSP-AG, 6/15/14<br> 
| <br> 
| 6/15/14 PHP<br>
|-
| 8<br> 
| PHP-DTBP-AG, 6/15/14<br> 
| <br> 
| 6/15/14 PHP<br>
|-
| 9<br> 
| PHP-DSP-AG, 7/17/14<br> 
| <br> 
| 7/17/14 PHP <br>
|-
| 10<br> 
| PHP-DTBP-AG, 7/17/14<br> 
| <br> 
| 7/17/14 PHP
|-
| 11<br> 
| Stability #1<br> 
| Store at RT<br> 
| <br>
|-
| 12<br> 
| Stability #1&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; <br> 
| Heat 50 oC<br> 
| <br>
|-
| 13<br> 
| Stability #2<br> 
| Store at RT 
| <br>
|-
| 14<br> 
| Stability #2 
| 50 oC&nbsp;&nbsp; <br> 
| <br>
|-
| 15<br> 
| Stability #3 
| Store at RT 
| <br>
|-
| 16<br> 
| Stability #3 
| 50 oC 
| <br>
|-
| 17<br> 
| Stability #4 
| Store at RT 
| <br>
|-
| 18<br> 
| Stability #4 
| 50 oC 
| <br>
|-
| 19<br> 
| Stability #5 
| Store at RT 
| <br>
|-
| 20<br> 
| Stability #5 
| 50 oC 
| <br>
|-
| 21<br> 
| Stability #6 &nbsp; <br> 
| Store at RT 
| <br>
|-
| 22<br> 
| Stability #6 
| 50 oC 
| <br>
|-
| 23<br> 
| Stability #7 
| Store at RT 
| <br>
|-
| 24<br> 
| Stability #7 
| 50 oC 
| <br>
|}

Picture of 8/2/14<br> 

[[Image:PHP-AG and stability 8 2 14 1.JPG]]<br> 

8/3/14 

[[Image:PHP-AG and stability 8 3 14.JPG]] 

8/4/14 

[[Image:PHP-AG and stability 8 3 14.JPG]] 

8/5/14<br>

<br>

[[Image:PHP-AG_and_stability_8_5_14.JPG]]<br>

<br>

8/3/14<br>

{| cellspacing="1" cellpadding="1" border="1" width="517" height="277"
|-
| Well<br>
| Sample<br>
| <br>
| <br>
|-
| 1<br>
| Stability #2<br>
| Store at RT<br>
| <br>
|-
| 2<br>
| Stability #2<br>
| 50 oC<br>
| <br>
|-
| 3<br>
| Stability #8<br>
| Store at RT<br>
| <br>
|-
| 4<br>
| Stability #8<br>
| 50 oC<br>
| <br>
|-
| 5<br>
| 7/17/14 PHP conjugated to PEI18 with DSP<br>
| 1:10<br>
| <br>
|-
| 6<br>
| 7/17/14 PHP conjugated to PEI18 with DSP<br>
| 1:30<br>
| <br>
|-
| 7<br>
| 7/17/14 PHP conjugated to PEI18 with DSP<br>
| 1:10<br>
| <br>
|-
| 8<br>
| 7/17/14 PHP conjugated to PEI18 with DSP<br>
| 1:30<br>
| <br>
|-
| 9<br>
| Stability #1<br>
| Store at RT<br>
| <br>
|}

Picture of 8/5/14<br>

[[Image:PHP-AG_and_stability_8_5_14_1.JPG]]<br>

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