User:Ajeffs/Notebook/Melanoma/2008/05/19: Difference between revisions

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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span>
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Kaloyan's melanoma project</span>
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
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==Kal's mini-project==
==The effect of PAX3 and MITF knockdown in melanoma cell lines==
===siRNA-mediated knockdown of PAX3 and MITF in NZM12 and NZM15===
===siRNA-mediated knockdown of PAX3 and MITF in NZM12 and NZM15===
*Reverse transfection with Dharmacon Smartpools (10 nM MITF; 20 nM PAX3; 20 nM non-targeting pool).
*Reverse transfection with Dharmacon Smartpools (10 nM MITF; 20 nM PAX3; 20 nM non-targeting pool).

Revision as of 17:24, 18 May 2008

Kaloyan's melanoma project <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

The effect of PAX3 and MITF knockdown in melanoma cell lines

siRNA-mediated knockdown of PAX3 and MITF in NZM12 and NZM15

  • Reverse transfection with Dharmacon Smartpools (10 nM MITF; 20 nM PAX3; 20 nM non-targeting pool).
    • Assess transcript knock-down at 24 hours post-txn by using qPCR
    • Assess protein levels 48 hours post-txn by using Western
    • n = 2 of 3 replicate experiments.
  • Measure EDNRB, RAB27A, FOXD3, and SNAI2 transcript levels by using qPCR at 24 and 48 hours post-txn.

Does miR-137 repeat expansion correlate with MITF levels in NZM cells?

  • Assess miR-137 locus stability at 1p22 with conventional PCR.
    • purify and sequence a selection of the different sized amplicons.
    • predict impact of repeat expansion on pre-miR folding.
    • measure miR-137 VNTR status in non-melanoma/skin-derived cells?


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