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Old 02-05-2016, 12:33 AM   #1
Ghada
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Location: Egypt

Join Date: Nov 2014
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Default 3500 genetic analyzer low quality value

Dear professor
we have a new 3500 genetic analyzer that sanger/dye terminator from applied biosystem. All amplicon have e low to medium quality value the same site 75-85 bp including the control PGEM, however the seq standard is ok. please check attached files
Any comments
regards
Ghad
Attached Files
File Type: pdf SeqStd 1.pdf (614.3 KB, 10 views)
File Type: pdf Naglaa.pdf (160.8 KB, 6 views)
File Type: pdf PGEMT lab Hidi.pdf (598.6 KB, 5 views)
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Old 02-08-2016, 07:25 AM   #2
MU Core
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Location: Columbia, Missouri

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Ghada, you're observing the presence of "dye blobs" which is the result of poor removal of unincorporated dye terminators from the post-sequencing reaction. A common issue in dye terminator sequencing. You'll want to revisit your reaction clean-up method to see if changes can be made to optimize the method. Or, you may want to consider a different approach to reaction clean-up. Performing a google search with "removing dye blobs" will provide you with information on the topic.
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Old 02-10-2016, 09:40 PM   #3
Ghada
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thank you MU core for your help.
Regards
Ghada
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