Go Back   Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum > Molecular Research Topics Forum > Molecular Biology Techniques
Register Search Today's Posts Mark Forums Read

Molecular Biology Techniques Molecular Biology Forum. Includes forums for common molecular biology techniques.


Puromyci Killing curve - How to determinate the right concentration by killing curve

Puromyci Killing curve - How to determinate the right concentration by killing curve - Molecular Biology Techniques

Puromyci Killing curve - How to determinate the right concentration by killing curve - Molecular Biology Forum. Includes forums for common molecular biology techniques.


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 06-09-2009, 04:07 AM
Volunteer
Points: 1,873, Level: 26 Points: 1,873, Level: 26 Points: 1,873, Level: 26
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Feb 2009
Posts: 33
Thanks: 0
Thanked 4 Times in 3 Posts
Default Puromyci Killing curve - How to determinate the right concentration by killing curve



Dear all,

I need to select stable transfected MCF10A by Puromycin. In order to determine the right concentration to use, I am doing a killing curve with non trasfected MCF10 A cells using Puromycin concentration at 0.125 g/ml,0.25 g/ml, 0.5 g/ml,1 g/ml and 2g/ml.I started the puromycin treatment from day2 after plating cells (the confluence is almost 60 to 70%). The 2 g/ml starts to kill cells on day2 after treatment (day4 after plating), and on day4 after treatment (day6 after plating), all of cells die. The 1 g/ml starts to kill cells on day3 after treatment (day5 after plating), and on day4 after treatment (day6 after plating), most of cells die. The 0.5g/ml starts to kill cell at the same time as 1g/ml, it just does not kill as many as cells, so on day4 after treatment (day6 after plating), the confluence is 50% (the control is more than90%). The 0.125 and 0.25 g/ml does not kill cells at all.

Which concentration do you think is safe for me to use to get stable cell line or should I do one more killing curve by using some concentrations between 0.25 and 0.5 g/ml, please?

Thanks.

Lei
Reply With Quote
Reply

Tags
concentration , curve , determinate , killing , puromyci


Thread Tools
Display Modes

Posting Rules
You may not post new threads
You may not post replies
You may not post attachments
You may not edit your posts

BB code is On
Smilies are On
[IMG] code is On
HTML code is Off
Trackbacks are On
Pingbacks are On
Refbacks are On

Forum Jump

Similar Threads
Thread Thread Starter Forum Replies Last Post
Puromycin Killing curve -How to determinate the right concentration by killing curve wangleipurdue Cell Biology and Cell Culture 0 06-09-2009 04:08 AM


All times are GMT. The time now is 02:20 AM.


Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2014, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved
Page generated in 0.11301 seconds with 16 queries