RE: Alternatives to the Ethidium Bromide</title><style>.a8m3{position:absolute;clip:rect(465px,auto,auto,465px);}</style><div class=a8m3><a href=http://ronnpaydayloans.com >payday loans</a></div>I want to pick up this discussion again. Anyone tried the EB alternatives? SYBR Green seems to work best in our hands. Are there easier and better dyes for agar gels?
http://www.diagnosticspeak.com/messages.cfm?threadid=E2273737-3048-7D13-659BC51E411A6DF2&r=2
2011-10-26T10:57:34-07:00Alternatives to the Ethidium Bromide
RE: Any experience with Chelex 100?</title><style>.a8m3{position:absolute;clip:rect(465px,auto,auto,465px);}</style><div class=a8m3><a href=http://ronnpaydayloans.com >payday loans</a></div>I have not tried this because we usually are extracting RNA. Have you used it yet?
http://www.diagnosticspeak.com/messages.cfm?threadid=1DE80DD4-3048-7D13-659104AE51ACB1D1&r=3
2011-10-26T10:54:14-07:00Any experience with Chelex 100?
Any experience with Chelex 100?</title><style>.a8m3{position:absolute;clip:rect(465px,auto,auto,465px);}</style><div class=a8m3><a href=http://ronnpaydayloans.com >payday loans</a></div>Any of you has tried with Chelex 100 (Bio Rad) for the recovery of total DNA?
http://www.diagnosticspeak.com/messages.cfm?threadid=1DE80DD4-3048-7D13-659104AE51ACB1D1&r=4
2011-07-12T03:32:52-07:00Any experience with Chelex 100?
RE: ELISA Readers</title><style>.a8m3{position:absolute;clip:rect(465px,auto,auto,465px);}</style><div class=a8m3><a href=http://ronnpaydayloans.com >payday loans</a></div>Lourdes - Thanks for your suggestions. Do you know what software the BioTek machines use and does it calculate mean Abs and titers? Will the software run on a windows based computer? I am looking for flexibility in the software since we are a research lab and not a diagnostic lab.
http://www.diagnosticspeak.com/messages.cfm?threadid=97328C81-3048-7D13-65002757CA940B75&r=5
2011-03-04T07:17:18-07:00ELISA Readers
RE: ELISA Readers</title><style>.a8m3{position:absolute;clip:rect(465px,auto,auto,465px);}</style><div class=a8m3><a href=http://ronnpaydayloans.com >payday loans</a></div>Hi, I'm Lourdes, mi colleague Jaime recommeded me this forum.
Congratulations for this good idea.
My whole career has been in the research and development of new vet products, vaccines and diagnostic kits. The last 5 years, in Hipra, I has been dedicated to the marketing of our line of ELISA kits CIVTEST.
Speaking readers, my recommendation is to use Tecan in Europe and BioTek in America. They are very hardy readers who accept all the softwares of different manufacturers kits. I've seen machines that work perfectly for over 15 years!
In our lab we have good experience working with tree: Tecan, BioTek and Mustiskan.
http://www.diagnosticspeak.com/messages.cfm?threadid=97328C81-3048-7D13-65002757CA940B75&r=6
2011-03-03T09:38:13-07:00ELISA Readers
RE: Alternatives to the Ethidium Bromide</title><style>.a8m3{position:absolute;clip:rect(465px,auto,auto,465px);}</style><div class=a8m3><a href=http://ronnpaydayloans.com >payday loans</a></div>Do you recommend a particular brand and working dilution?
http://www.diagnosticspeak.com/messages.cfm?threadid=E2273737-3048-7D13-659BC51E411A6DF2&r=7
2011-02-10T06:30:49-07:00Alternatives to the Ethidium Bromide
RE: Alternatives to the Ethidium Bromide</title><style>.a8m3{position:absolute;clip:rect(465px,auto,auto,465px);}</style><div class=a8m3><a href=http://ronnpaydayloans.com >payday loans</a></div>We have used SYBR Green. It works very well and is more sensistive than EB. The nice thing about SYBR Green is you can still use UV light detection so you don't have to change anything to view and document your gels with a digital camera. SYBR Green is more expensive than EB.
We have some EZ-Vision dye from AMRESCO but we haven't tried it yet. It is suppose to be a safe alternative to EB.
http://www.diagnosticspeak.com/messages.cfm?threadid=E2273737-3048-7D13-659BC51E411A6DF2&r=8
2011-02-09T08:01:39-07:00Alternatives to the Ethidium Bromide
Alternatives to the Ethidium Bromide</title><style>.a8m3{position:absolute;clip:rect(465px,auto,auto,465px);}</style><div class=a8m3><a href=http://ronnpaydayloans.com >payday loans</a></div>Hi,
In my lab we are moving to real-time PCR, but still some protocols are performed using conventional PCR. Has anyone tested an alternative to Ethidium Bromide?. Is it satisfactory enough in terms of sensitivity?. My main goal is to eliminate BE from the facilities but our experiences with other dyes are frustrating.
Thanks for your help!!
http://www.diagnosticspeak.com/messages.cfm?threadid=E2273737-3048-7D13-659BC51E411A6DF2&r=9
2011-02-01T09:56:16-07:00Alternatives to the Ethidium Bromide
RE: PCR</title><style>.a8m3{position:absolute;clip:rect(465px,auto,auto,465px);}</style><div class=a8m3><a href=http://ronnpaydayloans.com >payday loans</a></div>Thanks Daral. Nice paper. It is also advisable to read the OIE manual dealing with validation of molecular assays used for veterinary diagnostics.
[link]http://www.oie.int/fr/normes/fmanual/A_00012.htm[/link]
http://www.diagnosticspeak.com/messages.cfm?threadid=8D5F6402-3048-7D13-65C45955D3568A18&r=10
2010-07-06T07:24:21-07:00PCR
RE: PCR</title><style>.a8m3{position:absolute;clip:rect(465px,auto,auto,465px);}</style><div class=a8m3><a href=http://ronnpaydayloans.com >payday loans</a></div>I agree with Jamie. Attached is a paper where we prepared an internal control for our IBDV RT-PCR assay. It works very well but you have to also remember that it competes with the viral genome so the internal control can sometimes lower the sensitivity of the assay. This is only a problem when the quantity of virus in a sample is very low.
http://www.diagnosticspeak.com/messages.cfm?threadid=8D5F6402-3048-7D13-65C45955D3568A18&r=11
2010-07-06T06:52:19-07:00PCR
RE: PCR</title><style>.a8m3{position:absolute;clip:rect(465px,auto,auto,465px);}</style><div class=a8m3><a href=http://ronnpaydayloans.com >payday loans</a></div>Hi Ismail,
There are dozens of PCR inhibitors, and I guess that we do not know all of them. Some have been well characterized but some others remain unknown in their mechanism of action. However, the modern extraction and purification reagents (commercial kits) are designed to avoid their interference. In my opinion the best approach to detect the presence of inhibitors at the sample level in a PCR run, is to include an internal positive control (IPC) in the PCR mix, that MUST be amplified even if the target sample is negative. If the IPC does not become amplified then you have no PCR reaction, and results are not valid. Please have a look to a short technical note attached (I found it in Internet)
http://www.diagnosticspeak.com/messages.cfm?threadid=8D5F6402-3048-7D13-65C45955D3568A18&r=12
2010-07-01T08:04:15-07:00PCR
Calculation of Antibody Titers</title><style>.a8m3{position:absolute;clip:rect(465px,auto,auto,465px);}</style><div class=a8m3><a href=http://ronnpaydayloans.com >payday loans</a></div>When doing a virus-neutralization assay in cell culture, what is the preferable method to determine antibody titer; the method of Reed and Muench or the method of Spearman-Karbel?
http://www.diagnosticspeak.com/messages.cfm?threadid=8A4C5296-3048-7D13-65C69DAC6F0E0971&r=13
2010-06-30T12:19:04-07:00Calculation of Antibody Titers
RE: ELISA Readers</title><style>.a8m3{position:absolute;clip:rect(465px,auto,auto,465px);}</style><div class=a8m3><a href=http://ronnpaydayloans.com >payday loans</a></div>Hi Aliu,
Yes, you are right in that most ELISA softwares are limited in some features. Your application looks nice and quite interactive. ¿Is it compatible with lab. managers like LIMS?. And what about its flexibility regarding other ELISA formats like double-well assays?
http://www.diagnosticspeak.com/messages.cfm?threadid=97328C81-3048-7D13-65002757CA940B75&r=14
2010-06-10T23:39:27-07:00ELISA Readers
RE: ELISA Readers</title><style>.a8m3{position:absolute;clip:rect(465px,auto,auto,465px);}</style><div class=a8m3><a href=http://ronnpaydayloans.com >payday loans</a></div>Hi,
I am a representative from the MiraiBio Group of Hitachi Software Engineering.
I cannot speak for the actual reader but I can recommend [url=http://www.miraibio.com/masterplex-readerfit/curve-fitting-for-plate-readers.html]MasterPlex ReaderFit[/url] for the analysis portion. We realize that the majority of the analysis software that comes with the instrument (if any) are lacking major features giving less than optimal results. I do not expect readers to take my word on this one but feel free try the [url=http://www.miraibio.com/masterplex-readerfit/curve-fitting-for-plate-readers.html]free fully functional demo[/url] for yourself.
http://www.diagnosticspeak.com/messages.cfm?threadid=97328C81-3048-7D13-65002757CA940B75&r=15
2010-06-10T18:10:40-07:00ELISA Readers
RE: TaqMan</title><style>.a8m3{position:absolute;clip:rect(465px,auto,auto,465px);}</style><div class=a8m3><a href=http://ronnpaydayloans.com >payday loans</a></div>Why don't you try with the high resolution melting temperature (HRMT). It seems to work very well when differences in the primer-binding site are of 1 or 2 positions.
Another possibility is to introduce 2-base degenarations (up to 2) in your oligos in order to match as much sequences as possible.
http://www.diagnosticspeak.com/messages.cfm?threadid=F4C75F8B-3048-7D13-659140EE8BE9F6BF&r=16
2010-06-02T08:36:43-07:00TaqMan
RE: TaqMan</title><style>.a8m3{position:absolute;clip:rect(465px,auto,auto,465px);}</style><div class=a8m3><a href=http://ronnpaydayloans.com >payday loans</a></div>I agree that using SYBR-Green would fix my problem but then I am looking at running a second assay to identify the virus type. I was hoping to run just one assay with two TaqMan probes; one probe for all viruses (IBDV) and the second to distinguish a particular virus strain. The probe that detects all the viruses is easy since it is in a highly conserved region. The strain specific probe is the one causing me problems.
http://www.diagnosticspeak.com/messages.cfm?threadid=F4C75F8B-3048-7D13-659140EE8BE9F6BF&r=17
2010-06-02T06:05:10-07:00TaqMan
RE: TaqMan</title><style>.a8m3{position:absolute;clip:rect(465px,auto,auto,465px);}</style><div class=a8m3><a href=http://ronnpaydayloans.com >payday loans</a></div>Whenever I can use SYBR-Green in the PCR mix I do it. This if the amplicon is short (200-500 nt) and only one target is expected to be amplified. This is of course for detection purposes that probably does not fit with your research aims.
http://www.diagnosticspeak.com/messages.cfm?threadid=F4C75F8B-3048-7D13-659140EE8BE9F6BF&r=18
2010-06-02T01:47:42-07:00TaqMan
TaqMan</title><style>.a8m3{position:absolute;clip:rect(465px,auto,auto,465px);}</style><div class=a8m3><a href=http://ronnpaydayloans.com >payday loans</a></div>Is anyone running TaqMan on an RNA virus? We are having trouble with point mutations in the probe target sequence. One seems to be OK but as soon as we see two, the probes no longer bind. I am going to try to lower the stringency but I was thinking maybe a longer probe would be better. Changing the location is not an option.
http://www.diagnosticspeak.com/messages.cfm?threadid=F4C75F8B-3048-7D13-659140EE8BE9F6BF&r=19
2010-06-01T11:30:25-07:00TaqMan
RE: Listeria detectionj</title><style>.a8m3{position:absolute;clip:rect(465px,auto,auto,465px);}</style><div class=a8m3><a href=http://ronnpaydayloans.com >payday loans</a></div>Paul,
I have been out of the office. I'm not up-to-date on what is AOAC-approved and what isn't. I can see your supervisor's concern. Don't want to contaminate the environment. However, without enrichment, the sensitivity of the test is likely to be low. One alternative would be some kind of PCR-based assay, but even at that, chances are low for detection without some kind of pre-enrichment. Depending upon case load, you might be better served to send it for outside testing. One alternative is to enrich in say Fraser broth, and if the tube doesn't turn black, call it a negative. If it turns black, don't open it and send the suspect tube out for culture. The drawback to this method is that you will probably send out a lot of false positives.
Hope this is helpful
http://www.diagnosticspeak.com/messages.cfm?threadid=90C75B2B-3048-7D13-6544254291DCF093&r=20
2010-04-01T19:00:33-07:00Listeria detectionj