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FLP VLP Optimization 2014 3 4 - Revision history
2026-10-18T05:33:35Z
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https://eln.excellgen.com/index.php?title=FLP_VLP_Optimization_2014_3_4&diff=11426&oldid=prev
Paul.xu at 22:37, 8 March 2014
2014-03-08T22:37:04Z
<p></p>
<p><b>New page</b></p><div><br> <br />
<br />
= Making VLP<br> =<br />
<br />
== Co-transfect FRT reporter plasmid&nbsp;2013 with VSVG and FLP constructs<br> ==<br />
<br />
293T cells, 12 well plate<br> <br />
<br />
FRT plasmid: 2013 pCMV-FRT-TurboRFP-ODC-2Stops-FRT-TurboGFP, 0.3 ug<br> <br />
<br />
FLP plasmid: 0.6 ug<br> <br />
<br />
pCMV-VSVG (505): 0.6 ug<br> <br />
<br />
<br> <br />
<br />
{| width="583" border="1" cellspacing="1" cellpadding="1"<br />
|-<br />
| 2103 pCMV-RSV-Gag-Flpo, #1<br> <br />
| 2103 pCMV-RSV-Gag-Flpo, #2<br> <br />
| 2103 pCMV-RSV-Gag-Flpo, #3<br> <br />
| 2103 pCMV-RSV-Gag-Flpo, #1 maxi<br><br />
|-<br />
| 1969 pCMV5-Gag-p2b-NLS-Flp-WPRE-oriP<br> <br />
| 1962 pCMV5-HTN-Flpo<br> <br />
| 1445 pCMV5-Gag-NLS-Flp-WPRE-oriP<br> <br />
| 1981 pCMV5-Tat-Flp-PDGFR<br><br />
|-<br />
| 1964 pCMV5-n-NA-NLS-Flp-WPRE-oriP<br> <br />
| 1526 pCDNA3.1-NLS-FLPo <br> <br />
| 1525 pCDNA3.1-FLPe<br> <br />
| GFP contro, no 2013<br />
|}<br />
<br />
<br> <br />
<br />
[[Image:Flp VSVG 2014 3 4.JPG]]<br> <br />
<br />
<br> <br />
<br />
== Co-transfect FRT reporter plasmid 2013 with FLP constructs (no VSVG) ==<br />
<br />
293T cells, 12 well plate <br />
<br />
FRT plasmid: 2013 pCMV-FRT-TurboRFP-ODC-2Stops-FRT-TurboGFP, 0.5 ug <br />
<br />
FLP plasmid: 1.1 ug<br> <br />
<br />
{| width="622" border="1" cellspacing="1" cellpadding="1"<br />
|-<br />
| 2103 pCMV-RSV-Gag-Flpo, #1<br> <br />
| 2103 pCMV-RSV-Gag-Flpo, #2<br> <br />
| 2103 pCMV-RSV-Gag-Flpo, #3<br> <br />
| 2103 pCMV-RSV-Gag-Flpo, #1 maxi<br><br />
|-<br />
| 1969 pCMV5-Gag-p2b-NLS-Flp-WPRE-oriP<br> <br />
| 1962 pCMV5-HTN-Flpo<br> <br />
| 1445 pCMV5-Gag-NLS-Flp-WPRE-oriP<br> <br />
| 1981 pCMV5-Tat-Flp-PDGFR<br><br />
|-<br />
| 1964 pCMV5-n-NA-NLS-Flp-WPRE-oriP<br> <br />
| 1526 pCDNA3.1-NLS-FLPo <br> <br />
| 1525 pCDNA3.1-FLPe(cell death due to contamination) <br> <br />
| 2013&nbsp;pCMV-FRT-TurboRFP-ODC-2Stops-FRT-TurboGFP<br />
|}<br />
<br />
<br> [[Image:Flp No VSVG 2014 3 4.JPG]]<br> <br />
<br />
<br> <br />
<br />
= Activity assay<br> =<br />
<br />
add 0.5 ml condition culture medium to 24 well plate, 293T cells transfected with 2013 pCMV-FRT-TurboRFP-ODC-2Stops-FRT-TurboGFP<br> <br />
<br />
<br> <br />
<br />
== VSVG-FLP<br> ==<br />
<br />
[[Image:FLP VLP assay VSVG 2014 3 5.JPG]]<br> <br />
<br />
{| width="583" border="1" cellspacing="1" cellpadding="1"<br />
|-<br />
| 2103 pCMV-RSV-Gag-Flpo, #1<br> <br />
| 2103 pCMV-RSV-Gag-Flpo, #2<br> <br />
| 2103 pCMV-RSV-Gag-Flpo, #3<br> <br />
| 2103 pCMV-RSV-Gag-Flpo, #1 maxi<br><br />
|-<br />
| 1969 pCMV5-Gag-p2b-NLS-Flp-WPRE-oriP<br> <br />
| 1962 pCMV5-HTN-Flpo<br> <br />
| 1445 pCMV5-Gag-NLS-Flp-WPRE-oriP<br> <br />
| 1981 pCMV5-Tat-Flp-PDGFR<br><br />
|-<br />
| 1964 pCMV5-n-NA-NLS-Flp-WPRE-oriP<br> <br />
| 1526 pCDNA3.1-NLS-FLPo <br> <br />
| 1525 pCDNA3.1-FLPe<br> <br />
| GFP contro, no 2013<br />
|}<br />
<br />
== FLP (no VSVG)<br> ==<br />
<br />
[[Image:FLP VLP assay no VSVG 2014 3 5.JPG]]<br> <br />
<br />
{| width="622" border="1" cellspacing="1" cellpadding="1"<br />
|-<br />
| 2103 pCMV-RSV-Gag-Flpo, #1<br> <br />
| 2103 pCMV-RSV-Gag-Flpo, #2<br> <br />
| 2103 pCMV-RSV-Gag-Flpo, #3<br> <br />
| 2103 pCMV-RSV-Gag-Flpo, #1 maxi<br><br />
|-<br />
| 1969 pCMV5-Gag-p2b-NLS-Flp-WPRE-oriP<br> <br />
| 1962 pCMV5-HTN-Flpo<br> <br />
| 1445 pCMV5-Gag-NLS-Flp-WPRE-oriP<br> <br />
| 1981 pCMV5-Tat-Flp-PDGFR<br><br />
|-<br />
| 1964 pCMV5-n-NA-NLS-Flp-WPRE-oriP<br> <br />
| 1526 pCDNA3.1-NLS-FLPo <br> <br />
| 1525 pCDNA3.1-FLPe<br> <br />
| 2013&nbsp;pCMV-FRT-TurboRFP-ODC-2Stops-FRT-TurboGFP<br />
|}<br />
<br />
<br> [[Category:VLP]]</div>
Paul.xu