Biomod/2012/HKBU/BU Magician:Labook:Notebook/Labbook/2012/10/06

From OpenWetWare
Jump to navigationJump to search

<html> <style> .container{background-color: #FFFFFF; margin-top:0px}

  1. column-content {width: 0px; float: left; margin: 0 0 0 0;padding: 0;}

.firstHeading {display:none; width:0px;}

  1. column-one {display:none; width:0px;background-color: #FFFFFF;}
  1. globalWrapper{width: 920px; background-color: #FFFFFF; margin-left: auto;margin-right: auto;

-moz-box-shadow: 0px 0px 15px #ccc; -webkit-box-shadow: 0px 0px 15px #ccc; box-shadow: 0px 0px 15px #ccc;}

  1. content{ margin: 0 0 0 0; align: center; padding: 12px 12px 12px 12px; width: 800px;background-color: #FFFFFF; border: 0;}
  2. bodyContent{ width: 800px; align: center; background-color: #FFFFFF;}
  3. column-content{width: 800px;background-color: #FFFFFF;}

</style> <a href="">edit</a> </html> <html xmlns="" xml:lang="en" lang="en"> <head> <meta http-equiv="Content-Type" content="text/html; charset=UTF-8" /> <title>jOuery</title> <link rel="stylesheet" href="style.css" /> <style> .menu { height: 45px; display: block; } .menu ul { list-style: none; padding: 0; margin: 0; } .menu ul li { float: left; overflow: hidden; position: relative; text-align: center; line-height: 45px; } .menu ul li a { position: relative; display: block; width: 128px; height: 45px; font-family: Arial; font-size: 11px; font-weight: bold; letter-spacing: 1px; text-transform: uppercase; text-decoration: none; cursor: pointer; } .menu ul li a span { position: absolute; left: 0; width: 128px; } .menu ul li a span.out { top: 0px; } .menu ul li a span.over, .menu ul li a { top: -45px; }

  1. menu2 { background:#45A8DF; }
  2. menu2 ul li a { color:#FFF; }
  3. menu2 ul li a span.over { background: #A6DD00; color:#333; }
  4. menu2 ul li.nav1 a span.over { background: #fea274; }
  5. menu2 ul li.nav2 a span.over { background: #b0bbba; }
  6. menu2 ul li.nav3 a span.over { background: #a3f091; }
  7. menu2 ul li.nav4 a span.over { background: #86dbf9; }
  8. menu2 ul li.nav5 a span.over { background: #e0caf0; }
  9. menu2 ul li.nav6 a span.over { background: #9dace9; }
  10. menu2 ul li.nav7 a span.over { background: #FFE66F; }

</style> <script type="text/javascript" src=""></script> <script language="javascript"> $(document).ready(function() {

$("#menu2 li a").wrapInner( '<span class="out"></span>' );

$("#menu2 li a").each(function() { $('<span class="over">' + $(this).text() + '</span>' ).appendTo( this ); });

$("#menu2 li a").hover(function() { $(".out",this).stop().animate({'top':'45px'},200); $(".over",this).stop().animate({'top':'0px'},200);

}, function() { $(".out",this).stop().animate({'top':'0px'},200); $(".over",this).stop().animate({'top':'-45px'},200); });


</script> </head> <body> <div id="HEADER">

 <div id="menu2" class="menu" style="width: 897px">
     <li class="nav1"><a href="">Home</a></li>
     <li class="nav2"><a href="">Projects</a></li>
     <li class="nav3"><a href="">ResultS</a></li>
      <li class="nav7"><a href="">Methods</a></li>
     <li class="nav4"><a href="">Team</a></li>
     <li class="nav5"><a href="">Labook</a></li>
     <li class="nav6"><a href="">Extra</a></li>

</div> </body> </html>


According to the confocal result, we found that Plan A doesn’t work well. Therefore, we decided to use Plan B only.

Solution preparation

(1) Nanoparticle solution preparation
250μl of nanoparticle stock solution (conc.=40mg/ml) is diluted with 1750μl HBSS (Concentration: 5mg/ml)
Solution with concentration 5 times as working concentration is prepared here.
Total volume 500μl for each concentration

(2) SiRNA containing solution preparation:<br/> For each well, 300μl of nanoparticle solution is needed for treatment.
For SiRNA containing nanoparticle solution
60μl is added to 240 μl serum free medium to dilute 5 times.
Total volume: 60x 2(duplicates) x2(with magnetic field/without magnetic field)=240μl
Take in account the pipetting error, 240μl -> 320μl for each concentration solution containing nanoparticles

SiRNA stock concentration:40μM
SiRNA concentration needed for treatment 200nM
SiRNA concentration for nanoparticle solution preparation: 1000nM
( *Remarks:The final concentration of SiRNA containing nanoparticle solution is not calculated here.)
40*1000*X=1000(concentration)*320 X=8
Remarks: if under this condition, the nanoparticle is too concentrated for conjugation.

For SiRNA containing incomplete medium solution (experiment 9,10)
Volume of SiRNA containing medium
= 60μlx2 (duplicates) x 2(experiment 9,10)

Take in account of pipetting error, 240μl-> 320μl
After adding SiRNA to both nanoparticle and HBSS solution, the tubesare stored in 4 centigrade refrigerator to allow conjugating overnight for 24hrs.

Cell Seeding

(1)Precoat the 4 well plate
(2) Seeding cells
After cell count, calculate the volume should be added into each well according to the cell number of 5x104 cells/well.
Place in incubator overnight to allow cell growth