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Paul.xu at 21:30, 8 September 2014
2014-09-08T21:30:42Z
<p></p>
<p><b>New page</b></p><div>== Synthesis ==<br />
<br />
9/2/2014<br> <br />
<br />
1,2-Bis(3-aminopropylamino)ethane, APED<br>&nbsp; [[Image:1,2-Bis(3-aminopropylamino)ethane.png]]<br> http://www.sigmaaldrich.com/catalog/product/aldrich/239399<br>239399-100ML, $29.30<br> <br />
<br />
<br> <br />
<br />
{| width="255" cellspacing="1" cellpadding="1" border="1"<br />
|-<br />
| <br> <br />
| DBP<br> <br />
| DAP<br> <br />
| APED<br> <br />
| MeOH<br><br />
|-<br />
| D1<br> <br />
| 61 mg<br> <br />
| 12.5 mg<br> <br />
| 5 ul<br> <br />
| 75 ul<br><br />
|-<br />
| D2<br> <br />
| 61<br> <br />
| 12.5&nbsp;&nbsp;&nbsp; <br> <br />
| 10<br> <br />
| 75<br><br />
|-<br />
| D3<br> <br />
| 61<br> <br />
| 12.5 <br />
| 15<br> <br />
| 75<br><br />
|-<br />
| D4<br> <br />
| 61<br> <br />
| 12.5 <br />
| 20<br> <br />
| 75<br><br />
|-<br />
| D5<br> <br />
| 61<br> <br />
| 12.5 <br />
| 25<br> <br />
| 75<br><br />
|-<br />
| D6<br> <br />
| 61<br> <br />
| 12.5 <br />
| 30<br> <br />
| 75<br><br />
|}<br />
<br />
366 mg DBP + 75 mg DAP+ 150 ul MeOH--&gt;335 ul<br> <br />
<br />
Divide into 6 vials<br> <br />
<br />
+ 5 to 30 ul APED <br> <br />
<br />
+ 75 ul MeOH<br> <br />
<br />
50 <sup>o</sup>C react o/n<br> <br />
<br />
then 60 <sup>o</sup>C react 3 hours.<br> <br />
<br />
Purify with 0.5 ml isopropanol, 2X<br> <br />
<br />
Dissolve in 500 ul H2O<br> <br />
<br />
== Transfection ==<br />
<br />
9/4/14 Transfection: dilute 1:10, 1:30, 1:50, 1:75, use 3 ul + 1 ug DNA to transfect 293T cells in 24 well plate<br> <br />
<br />
9/5/14 Morning<br> <br />
<br />
[[Image:PHP-APED 2014 9 5 1.JPG]]<br> <br />
<br />
9/5/14 Afternoon<br> <br />
<br />
[[Image:PHP-APED 2014 9 5 2.JPG]]<br> <br />
<br />
9/6/14 Morning<br> <br />
<br />
[[Image:PHP-APED 2014 9 6 .JPG]]<br> <br />
<br />
9/7/14 <br />
<br />
[[Image:PHP-APED 2014 9 7.JPG]] <br />
<br />
9/8/14<br />
<br />
[[Image:PHP-APED_2014_9_8.JPG]]<br />
<br />
<br> <br />
<br />
== Comparison of TREN1, PHP-TREN, PHP-s-s-TREN and PHP-APED<br> ==<br />
<br />
TREN1, PHP-TREN (A2, A3, B1): see this page: [[Synthesis of PHP-TREN|Synthesis_of_PHP-TREN]]<br> <br />
<br />
PHP-TREN (C1): [[Synthesis of PHP-s-s-TREN|Synthesis_of_PHP-s-s-TREN]]<br> <br />
<br />
PHP-APED (D6)<br> <br />
<br />
=== Transfection ===<br />
<br />
9/5/14 <br />
<br />
Dilution: 1:10, 10:30, 1:50, 1:75 <br />
<br />
DNA 1 ug <br />
<br />
293 cells in 24 wells <br />
<br />
<br> <br />
<br />
=== Result ===<br />
<br />
9/6/14: <br />
<br />
Dead cells <br />
<br />
<br> <br />
<br />
{| width="200" cellspacing="1" cellpadding="1" border="1"<br />
|-<br />
| <br> <br />
| TREN1 <br />
| C1 <br />
| D6 <br />
| A2 <br />
| A3 <br />
| B1<br />
|-<br />
| 1:10 <br />
| 99% <br />
| 95% <br />
| 50% <br />
| 25% <br />
| 40% <br />
| 20%<br />
|-<br />
| 1:30 <br />
| 99% <br />
| 10% <br />
| 2% <br />
| 15% <br />
| 15% <br />
| 1%<br />
|-<br />
| 1:50 <br />
| 95% <br />
| 30% <br />
| 20% <br />
| 15% <br />
| 15% <br />
| 1%<br />
|-<br />
| 1:75 <br />
| 80% <br />
| 5% <br />
| 40% <br />
| 15% <br />
| 2% <br />
| 1%<br />
|}<br />
<br />
9/6/14 Morning <br />
<br />
[[Image:PHP-APED PHP TREN 2014 9 6.JPG]]<br> <br />
<br />
9/6/14 Afternoon <br />
<br />
[[Image:PHP-APED PHP TREN 2014 9 6 2.JPG]] <br />
<br />
9/7/2014 <br />
<br />
[[Image:PHP-APED PHP TREN 2014 9 7.JPG]] <br />
<br />
9/8/14<br />
<br />
[[Image:PHP-APED_PHP_TREN_2014_9_8.JPG]]<br />
<br />
[[Category:Paul]] [[Category:Transfection]]</div>
Paul.xu