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FW: microsatellites problems ABI 310

FW: microsatellites problems ABI 310 - Protocols and Methods Forum

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  #1  
Old 12-15-2005, 03:29 PM
Deanne Bell
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Default FW: microsatellites problems ABI 310



This is just an idea - but I found that my 310 was very sensitive to
changes in ambient temperatures. Results would be - different molecular
weights when I ran over night vs during the day . . . I tried getting a
thermostat in the room that kept a constant temperature within 3-5
degrees F and that solved my problem. I know that there is a heating
plate for the capillary but . . . theoretical isn't always actual.

More of my 2 cents - How big are the fragments you are running? Do you
really need something 2500 bp long? I seem to recall an ABI standard
that went from 35 to 500 base pairs. Standards I bought from other
companies always had a lot of breakdown (minor noise around the 'real'
band). AND if you let standards get too old (even ABIs), they will
start breaking down.

hope this helps
Deanne Bell
USDA Agricultural Research Service
Crop Diseases, Pests & Genetics
9611 South Riverbend Avenue
Parlier, CA 93648
voice (559) 596-2806
fax (559) 596-2897
[Only registered users see links. ]





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Old 04-19-2012, 01:55 PM
Pipette Filler
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Default Re: FW: microsatellites problems ABI 310

Hi,

I'm using the ABI 310 to run LOH analysis for the FH gene. I'm a new user of the ABI and I keep having a problem after I run my experiment. When I try to analyze my results a message "No sizing data" is display and I don't get any peaks. If I look at the internal size standard that I'm using (GS 500) I can see some peaks but not at the expected sizes.

If anyone could shed some light on this respect, I would really appreciate it

Thanks
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Old 04-27-2012, 01:41 PM
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Default Re: FW: microsatellites problems ABI 310

Are you using the Liz or the Rox size standard?

Have you looked at your raw data in genemapper? Are the peaks of the standard visible there? Could you post a screenshot of your traces? (or post an .fsa file)
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