GeneQuant-Summary of responses
John Hachey
HACHEY at EM.AGR.CA
Sat Jan 27 08:30:08 EST 1996
** High Priority **
Herewith are (most of) the responses I got to my enquiry regarding
people's
experience with the genequant spec. Seeing how it is about half
the price of a
full-blown spec and all we really would use a spec for is for
DNA/RNA, it may
be worth putting up with its annoying piccadillios.
P.S. All standard disclaimers apply to the folowing replies; and
thanks again
to all the respondees. -JH
P.P.S. I still haven't made up my mind.
-------------------------------
From: Alan Schoenfeld <aschoen at aecom.yu.edu>
(INTERNET:"aschoen at aecom.yu.edu")
To: DLETHRB.WLETHRB(HACHEY)
Date: January 11, 1996, (Thursday) 5:59am
Subject: (no subject)
HACHEY at EM.AGR.CA (John Hachey) wrote:
>Subject: Genequant II-comments?
>We are thinking of purchasing a GeneQuant II unit primarily for
>DNA and RNA quantitation.
>Can some owners of this machine share their reviews of this
>instrument?
Our lab has been using a Gene Quant II for a year and a half.
While the rest of the lab has no problem with it, I do. My
complaints:
1) Can only be used for DNA and RNA (260, 280 nm only; no visible)
2) I have found it to be unreliable when determining
concentrations
of plasmids, especially at concentrations above 1
microgram/microliter; sometimes I get a reading that is way off
for no apparent reason
3) Is awkward to use because you have to insert and remove the
cuvette at specific times when the machine beeps; if you're too
slow, you have to do it again (sometimes annoying)
It has been useful when quantitating oligos however and the nice
part about it is that if you enter the sequence, it figures out
the molarity (in picomoles) for you. This has been fairly
reliable.
I hope this helps.
Alan
--------------------------
From: Jennifer L. Potter <jras at post.its.mcw.edu>
(INTERNET:"jras at post.its.mcw.edu")
To: DLETHRB.WLETHRB(HACHEY)
Date: January 10, 1996, (Wednesday) 8:35pm
Subject: Re: Concealed
Dear John,
We have (I believe) the older version of the Gene Quant II, the
Gene Quant. We use it for both DNA and protein measurements. As
far as I know, no one in the lab uses the fancy calculator part of
the machine.
A few comments that may or may not be pertinent to the Gene Quant
II:
The machine is a single beam spec...and therefore has greater
variability at lower absorbances. For example, I blanked the
machine with water and then tested the exact same sample about 10
times. The A260 varied between -0.01 and +0.01. Our double beam
spec, however, does not deviate in this manner (I am told this is
an intrinsic difference between single and double beam specs).
Comparing highly absorbing samples (>1.0) with our double beam
showed the two readings to be identical.
As a personal preference, I really don't care for the way the
machine beeps at you to tell you when to insert and remove the
cuvette. If you're not alert and focused, you'll be too slow and
it will beep at you again and make you start all over. Pharmacia
people use this,however, as a selling point because it conserves
lamp time.
One other point, our manual did not describe all the different
functions and how to use them in great detail. I'd expect the
manual for the Gene Quant II to be better.
I'm sorry I can't fill you in on more detail about the calculator
function, etc. but hope I've given you some useful information.
We
use ours quite frequently and it has held up fine for the 2-3
years we've owned it. The only down side that I see is the
variability at low absorbances...before I actually tested the
same
sample 10 times in a row, I kept wondering where my DNA went! :)
Best of luck,
Jennifer L. Potter
Medical College of WI
Dept of Biochemistry
--------------------------------
From: sathish <sathish at biokemi.su.se>
(INTERNET:"sathish at biokemi.su.se")
To: John Hachey <HACHEY at EM.AGR.CA>
Date: January 10, 1996, (Wednesday) 2:58am
Subject: Re: Genequant II
Hello:
We have been using GeneQuant II from Pharmacia for about 7 or 8
months. We have mixed feelings about it.
It is good because:
1. We do not have to wait for the machine to warm up.
2. It does most of the calculations, such as picomols, etc.
3. It is especially useful for quantitating oligos.
4. People from neighbouring labs also use it.
Some of the negative aspects:
1. If you have contaminations such as polysaccharides (RNA
preps!),
silica (geneclean), etc, the readings may not be true.
2. The software is designed in a round-about fashion, i.e. one
has to push the setup, enter buttons a few times before being
able to select DNA/RNA/Oligo, dilution factor etc.
3. The printer is not included (they gave free printers for a
month, a month after we purchased it. We were not lucky to
get one free. They have still not answered us if we can
somehow plug it to our computer network and download the data,
either.)
4. Measuring O.D. from very little volumes is posible only if you
are ready to shell out some money for the cappillary cuvettes!
Hope this helps.
sathish http://grolsch.biokemi.su.se/sathish/sathish.html
P.S. GeneQuant is different from GeneQuant II
-----------------------------
From: Steve Stelman <steves at burn.ucsd.edu>
(INTERNET:"steves at burn.ucsd.edu")
To: John Hachey <HACHEY at EM.AGR.CA>
Date: January 9, 1996, (Tuesday) 1:50pm
Subject: Re: Concealed (Genequant II)
John Hachey wrote:
> > Subject: Genequant II-comments?
> We are thinking of purchasing a GeneQuant II unit primarily for
> DNA and RNA quantitation.
> Can some owners of this machine share their reviews of this
> instrument?
> Thanks, John.
In short, here are a few of the pro's and con's
Pros:
-more accurate quantitation than using a spec
-easier than our fluorometer, which is difficult to zero
-light source does not need to warm-up before use
-calculates concentrations based on A260 and A280 in single read
-has many calculation functions built in
Cons:
-cuvette not included (expensive to purchase from them)
-cuvette must be inserted and removed on cue from machine
within a few seconds, or reading is aborted. This
gets to be REALLY annoying, as you have to hover
over the machine with the cuvette waiting for the
beep.
Nevertheless, I use this machine regularly as opposed to the
alternatives.
-- Steve Stelman steves at burn.ucsd.edu
------------------------
From: Beverly Steele Allen, Ph.D. <ALLENB at moffitt.usf.edu>
(INTERNET:"ALLENB at moffitt.usf.edu")
To: John Hachey <HACHEY at em.agr.ca>
Date: January 9, 1996, (Tuesday) 11:21am
Subject: Re: Concealed
We have one as a piece of core equipment (2-3 labs use it) and so
far, we all love it. It's basically a narrow-range
spectrophotometer with a fancy calculator in it; doesn't take up
as much bench space as a full-size spec, and calculates your
recovery or whatever for you.
Good luck,
Beverly
__
|| Beverly S. Allen
==== Bone Marrow Transplant Program
| |__ University of South Florida
| |-.\ 12901 Bruce B. Downs Blvd., MDC 19
|__| \\ Tampa, FL 33612 (813)979-3960
|| || allenb at moffitt.usf.edu
=====__| My .sig is a .glock
________||__ Grand piano + mineshaft = C# or Bbmin
/___<><______\
-----------------------
From: Beverly Steele Allen, Ph.D. <ALLENB at moffitt.usf.edu>
(INTERNET:"ALLENB at moffitt.usf.edu")
To: DLETHRB.WLETHRB(hachey)
Date: January 9, 1996, (Tuesday) 11:33am
Subject: Genequant
PS.-Saw a posted reply about the genequant being quirky--so was
ours, until we plugged it into a surge-suppressor. It was giving
us really wierd and irreproducible readings one day while plugged
into the recepticle that was running a thermal cycler. Noticed
that when the thermal cycler kicked on, the genequant went
squirrelly. No problems with reproducibility after putting the
genequant plug into the computer's surge suppressor outlet.
No connection with whomever sells the thing!
Beverly
__
|| Beverly S. Allen
==== Bone Marrow Transplant Program
| |__ University of South Florida
| |-.\ 12901 Bruce B. Downs Blvd., MDC 19
|__| \\ Tampa, FL 33612 (813)979-3960
|| || allenb at moffitt.usf.edu
=====__| My .sig is a .glock
________||__ Grand piano + mineshaft = C# or Bbmin
/___<><______\
--------------------------
From: <Jfloring at aol.com> (INTERNET:"Jfloring at aol.com")
To: DLETHRB.WLETHRB(HACHEY),GW1.INTERNET("methods-and-
Date: January 9, 1996, (Tuesday) 10:36am
Subject: Re: Genequant
Re: Genequant: Ours was covered with graffiti within a month,
and
despised.
We did use it, but only when it wasn't important. It's
inaccurate, and liable to freakouts. You can't really control
anything. Arghhhhhhhh!!! The memory is painful! I use the spec,
now, for everything.
Cheers, Jeanne (happily genequantless)
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