Reduction plaque assay : carboxy methyl cellulose

J. Martinez-Irujo jjmirujo at unav.es
Mon Oct 25 05:02:46 EST 1999


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>To prepare the solution add 75 ml of hot Hank's Balanced Salt Solution
to  >an appropriate amount of CMC (we used 3.5 g, but probably you
should add >less CMC ), dissolve it and autoclave the solution. In
sterile conditions mix: >20 ml of this solution (cold), 400 ml of DMEM
and antibiotic.  Mix well and >store at 4 C. Add serum and glutamine
just before adding this solution to the >cells.

Sorry, this recipe is misleading (the final concentration of CMC should
be 0.7%). The correct one is:

add 75 ml of hot Hank's Balanced Salt Solution to an appropriate amount
of CMC (we used 3.5 g, but probably you should add less CMC ), dissolve
it and autoclave the solution. In sterile conditions mix this solution
(cold) with 400 ml of DMEM, 20 ml of  Hank's Balanced Salt Solution and
antibiotic to give a final volume of 500 ml.  Mix well and store at 4 C.
Add serum and glutamine just before adding this solution to the cells.

--
Juan Jos=E9 Mart=EDnez Irujo

Departamento de Bioquimica
Edificio de Investigaci=F3n
Universidad de Navarra
31080 (Aptdo. 177)  Pamplona, Spain

Tel:  34-948-425600  ext. 6484
Fax:  34-948 425649
e-mail:  jjmirujo at unav.es


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<html>
<i>>To prepare the solution add 75 ml of hot Hank's Balanced Salt Solution
to&nbsp; >an appropriate amount of CMC (we used 3.5 g, but probably you
should add >less CMC ), dissolve it and autoclave the solution. In sterile
conditions mix: >20 ml of this solution (cold), 400 ml of DMEM and antibiotic.&nbsp;
Mix well and >store at 4 C. Add serum and glutamine just before adding
this solution to the >cells.</i><i></i>
<p>Sorry, this recipe is misleading (the final concentration of CMC should
be 0.7%). The correct one is:
<p>add 75 ml of hot Hank's Balanced Salt Solution to an appropriate amount
<br>of CMC (we used 3.5 g, but probably you should add less CMC ), dissolve
<br>it and autoclave the solution. In sterile conditions mix this solution
(cold) with 400 ml of DMEM, 20 ml of&nbsp; Hank's Balanced Salt Solution
and antibiotic to give a final volume of 500 ml.&nbsp; Mix well and store
at 4 C. Add serum and glutamine just before adding this solution to the
cells.
<p>--
<br>Juan Jos&eacute; Mart&iacute;nez Irujo
<p>Departamento de Bioquimica
<br>Edificio de Investigaci&oacute;n
<br>Universidad de Navarra
<br>31080 (Aptdo. 177)&nbsp; Pamplona, Spain
<p>Tel:&nbsp; 34-948-425600&nbsp; ext. 6484
<br>Fax:&nbsp; 34-948 425649
<br>e-mail:&nbsp; jjmirujo at unav.es
<br>&nbsp;</html>

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