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leelee

Member Since 09 Mar 2009
Offline Last Active May 23 2012 12:48 AM
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Posts I've Made

In Topic: Question about measuring bacterial OD

16 May 2012 - 12:51 AM

When you say you usually measure 1ml- is that in a cuvette in a spectro? If yes, then you aren't actually measuring the whole 1ml are you? Just the absorbance of light that passes through that small bit of the cuvette in the middle somewhere. Volume is irrelevant- its all about path length.

My guess is that the micro plate reader has been set up in such a way as to calculate the OD600 accurately for a 200ul volume. So when you add a different volume, you stuff the readings up. Again, as for spectro the volume is almost irrelevant- the 200ul is used so that the path length is known, and that is all.
So adding 100ul, or 300ul and you change the path length, making your readings useless.

(If I am understanding what you are doing correctly)

In Topic: Blood tube for WBC analysis

15 May 2012 - 05:22 AM

You can lyse them using a red blood cell lysis buffer? That is pretty common practice. You should be able to find a protocol and recipe fairly easy by asking Professor Google :)

In Topic: Help to optain a long PCR produc

11 May 2012 - 04:52 PM

What are your reaction conditions?

In Topic: 7500 Reaction Volume

10 May 2012 - 08:49 AM

I don't think you can. I thought that the volume was used to help determine the ramping times for each temp cycle of the reaction. So I think its too late, and the run would have been optimised for whatever volume you entered.

Whether this is a major problem or not I'm not sure. What was your volume and what did you set it at?

As for normalising your data, sorry I'm absolutely no help to you there...sorry. Hope someone else has some ideas!
(maybe comparing your standards to another run?)

In Topic: I'm desperately seeking for the program that I used to generate this alignm

10 May 2012 - 08:44 AM

I think it is CLC

You could always download the trial version to check if I'm right?

http://www.clcbio.com/index.php?id=16

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