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Synthesis PHP-CBA-DSP-PEI18 - Revision history
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Paul.xu at 14:43, 22 January 2014
2014-01-22T14:43:00Z
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<p><b>New page</b></p><div>== Synthesis of PHP ==<br />
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1.5 g solid 1,3-Diamino-2-propanol, MW&nbsp; 90.12, 16.6 mmole <br />
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+3 ml MeOH <br />
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+3.63 g 1,3-dibrono-2-propanol&nbsp;, MW&nbsp; 217.89, 16.7 mmole<br>50 oC for 48 hours <br />
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store at -20 oc. <br />
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add 5 ml H2O to dissolve <br />
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+ 25 ml EtOH, mix, spin 3000 rpm for 5 min <br />
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remove supernatant <br />
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Add 5 ml H2O, + 25 ml EtOH <br />
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spin, repeat 2 more times <br />
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yield: 2.8 g wet PHP + H2O to 10 ml <br />
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== Synthesis PHP-DSP-PEI<br> ==<br />
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3 ml PHP solution, + 25 mg PEI18, + 25 mg DSP, 40 oC reaction o/n <br />
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<br> <br />
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9-26-2011<br> <br />
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4.5 ml PHP solution, + 37 mg PEI18, + 37 mg DSP, 40 oC reaction o/n <br> <br />
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== Synthesis PHP-CBA-PEI ==<br />
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3 ml PHP solution + 25 mg CBA, + 25 mg PEI18, 40 oC reaction o/n <br />
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== Guanydinylation<br> ==<br />
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[[Image:Guanydinylation_1_22_14_1.JPG]]<br />
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[[Image:Guanydinylation_1_22_14_2.JPG]]<br />
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== Transfection ==<br />
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Make 1:10, 1:30, 1:40, 1:50, 1:75, 1:100 dilution <br />
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use 3 ul diluted polymer sol, + 1 ug DNA, 50 ul DMEM for 12 well plate. incubation time: 20 min <br />
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1: PHP-CBA-PEI, 1:10<br>2: PHP-CBA-PEI, 1:30<br>3: PHP-CBA-PEI, 1:40<br>4: PHP-CBA-PEI, 1:50<br>5: PHP-CBA-PEI, 1:75<br>6: PHP-CBA-PEI, 1:100<br>7: PHP-DSP-PEI, 1:10<br>8: PHP-DSP-PEI, 1:30<br>9: PHP-DSP-PEI, 1:40<br>10:PHP-DSP-PEI, 1:50<br>11: PHP-DSP-PEI, 1:75<br>12: PHP-DSP-PEI, 1:100 <br />
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== Results ==<br />
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=== 12 hour after transfection ===<br />
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[[Image:PHP-CBA-DSP-PEI conjugate transfection comparison 7 14 2011 1.jpg]] <br />
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=== Day 1 after transfection ===<br />
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[[Image:PHP-CBA-DSP-PEI conjugate transfection comparison 7 14 2011.jpg]] <br />
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=== Day 2 after transfection ===<br />
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[[Image:PHP-CBA-DSP-PEI conjugate transfection comparison 7 15 2011.jpg]] <br />
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=== Day 3 after transfection ===<br />
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[[Image:PHP-CBA-DSP-PEI conjugate transfection comparison 7 16 2011.jpg]] <br />
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<br> <br />
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[[Image:PHP-CBA-DSP-PEI conjugate transfection comparison 7 18 2011.jpg]] <br />
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July 17, 2011 <br />
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[[Category:Transfection]] [[Category:Paul]]</div>
Paul.xu