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Fluoresce

Member Since 09 Feb 2013
Offline Last Active Jun 08 2013 11:55 AM

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About Me

Religion is a myth created by man to trick you & I, to consume you & I. Been trying to dodge it. Just believe in good & do good, AVOID THE DARK SIDE!

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Active Posts 12
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My research interests
Fluorescence, DNA Purification, Quantification, Immunology, Cell Biology, HIV

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Latest Visitors

robinlee
05 Jun 2013 - 07:26
blaszcz
29 May 2013 - 12:04
pito
27 May 2013 - 10:35

Posts I've Made

In Topic: Cell Debris Isolation From Blood

08 June 2013 - 11:34 AM

Hi gfischer...
Thanks for your reply. No I was more thinking in the direction of filteration (tandem filteration) using micro filter polymers. But iodixanol seems to be a promising way as well. I need to be able to separet the blood particles from stressed blood and study them in a microfluic system (i.e spike certain amount of these debris into intact blood and run them in a microfluidic device to find the threshold for failure). Do you have a good protocol to suggest for using iodixanol for this purpose? I would really be very grateful if you could help me out with this.
Thanks!
FL.

In Topic: Sensitive, Consistent and Reproducible Method(s) for Extraction of gDNA from Low

01 June 2013 - 07:26 PM

Thanks a lot for your precise and detailed reply on my queston regarding extraction from low volume blood. Your protocol seems very promising. Thanks again!
FL

In Topic: Help: Best way to disrupt blood cells to generate blood debris and gDNA to be us

10 March 2013 - 06:40 PM

Awesome! Thanks so much for your help. I really appreciate it. I'm going to try the dry ice/isoproponaol/37 C bath...This defintely is going to happen first thing when I get my hands on some blood this week

Thanks again and have a great week ahead!

In Topic: Help: Best way to disrupt blood cells to generate blood debris and gDNA to be us

10 March 2013 - 11:58 AM

Hello Bob...There you go...I always get the best answers from you. Thanks a lot. How long would you keep the cycles going though?

In Topic: Qiagen column eluted DNA gives higher concetration than the original sample

12 February 2013 - 12:00 AM

Hi Carel, This is very nice of you to take the time to help. Great way to approcach the calculations. I think you are rihgt. I'm going to have to check a few things to make sure nothing is coming from contaminants low 260/280 ratio for example (However unlikely). I also need to run the gel, I am a lttle afraid of the conformational changes of larger DNA fragments (i.e gDNA samples). The fluorescent molecules access to DNA fragmetns (Specially the way PicoGreen binds to DNA) can also be a factor. I know two out of three samples I tested had issues with degradation. I still have to run the gel and check the bands.

My pipetted volumes were above 5 ul, it's unlileky that this is the issue. I agree on the elution volume collected from columns. It's hardly 100 ul, however it's usually around 90-95 ul. Thanks again for your help. I really appreciate it.

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