User:Ajeffs/Notebook/Melanoma/2008/05/19

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 * style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]] Kaloyan's melanoma project
 * style="background-color: #F2F2F2" align="center"|  |Main project page
 * style="background-color: #F2F2F2" align="center"|  |Main project page


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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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