Although I don't use radioactivity often, there are still a few
procedures where I do or would choose to use it.
For blotting/hybridization procedures, I would still choose
radioactivity, hands down. As far as I know, most non-radioactive
detection procedures require an immunoblot-like dete
My opinion is that you should ALWAYS sequence PCR products if in the
end it will be important to have it error free (i.e. you are trying
to express active protein). Even with a high fidelity enzyme, there
can still be an errors . Sequencing now is a lot easier than doing a
bunch of experime
As Peter suggested, if this is for recloning into another vector, it
may be easier to "shotgun" clone both fragments and sort them out
later since it is relatively easy to distinguish small differences in
fragment size, but sometimes difficult to cleanly isolate such
fragments.
If you pla
I'm not sure I understand what you are doing. Really, for recombinant
protein expression you only would need to recover a single colony
containing your construct. People typically grow liquid cultures to
produce recombinant proteins.
If you are including IPTG in your plates (and it sounds l
On Feb 2, 2010, at 6:16 AM, Nick Theodorakis wrote:
On Feb 2, 5:26 am, WS wrote:
Dear Experts,
is there any commercial (and preferably cheaper) alternative to PD-10
desalting columns?
(They are filled with Sephadex G-25, sample volume is up to 2.5ml,
elution volume is 3.5ml).
Buy the empt
Does anything grow from these on plates with no selection?
A conservative thing to try might be to remove a large chunk of the
frozen stock and grow it in LB medium without selection for an hour
or so... maybe this would give any viable cells a bit of time to
recover if they weren't froz
Wo, from your post, it's not quite clear exactly what type of product
you are talking about and what the application is. For plant tissue
culture, we use 3M micropore tape (a sort of porous cloth tape),
which when sold to be used in tissue culture can be quite expensive.
I buy it from an on
---
On Oct 24, 2011, at 10:02 AM, AllisonH wrote:
> On 24/10/2011 3:54 AM, DK wrote:
>> In article,
>> lautys wrote:
>>> APS should prepare fresh every time you do your gel.
>>
>> For how much longer will this myth be propagated?
>>
>> There is nothing wrong with aliquoting APS and storing
I've not done much comparison, but I've been using Bio-Rad's Immun-
Star substrate for years. I use it with PVDF. There's an added
enhancer for use with nitrocellulose, but I don't use NC, so I don't
now how well that works. I've found the signal very long lived.
Don't know about the price e
I think this is a needless worry! Water has almost no buffering capacity, so a
tiny amount of dissolved CO2 impacts water's pH. This amount of CO2 is very
small compared to the amount of ACTUAL buffer (TRIS, phosphate, whatever) you
are using, so will generally have only a negligible effect on p
What do you want to use the protein for?
I had a very dilute, yet viscous extract of flowers that I wanted to use for
western blotting. Carbohydrates made it visous, interfered with protein assays,
and made the samples run quite poorly in a gel. I ended up using a phenol
extraction method follo
This is one of my all time favorite from when I was in grad school:
http://www.history.ucsb.edu/faculty/marcuse/images/ComicGradSchoolHell72dpi550pxw.jpg
---
Michael L. Sullivan, PhD
Research Molecular Geneticist
US Dairy Forage Research Center
1925 Linden Drive
Madison, WI 53706
608-890-0046 (Pho
Well Ed can clarify. His original post is somewhat ambiguous: he didn't really
say what he wanted, just that he wanted to precipitate the RNA. It sounded to
me like he wanted to get rid of RNA from a plasmid miniprep, especially with
his follow up post. But I agree that if he wanted high qualit
We've had the 9700 (over 10 years) for quite some time (over 10 years) with no
real problems. Are you using the little black tube insert for single tubes and
the red one for strip tubes? Without those you will crush the tubes (but I
think these are similar to what gets used with the 2700 (which
Yeah, I haven't quite figured out why they designed it this way, although the
tube holder is sort of nice for working with the strip tubes. I don't think
there's a way to adjust how much pressure the lid applies (like the MJ DNA
engine [now biorad?] has), so maybe ABI's approach with the 9700 wa
used to extract the tissue, in addition there is
>> loss due to apparatus (such as homogenizers), etc. There should be some
>> non-enzymatic standard for normalization, such as the Bradford assay or
>> proteins. Even better look at a composite score such as observed over
>>
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