SeqLab E-mailing list provides fast way to get
information maintenance breaks, holidays, changes in protocols etc.
To join the list, send an e-mail to pekka.ellonen(ät)ktl.fi.
Your personal GIU account number is shown in
Sequencing Browser software when you login with your personal
username and password.
Click here for
help.
SeqLab application folder is visible for all SeqLab
customers in GIU environment. When you apply for GIU account you should at least select
"Med Genetics SeqLab" option in the form. If you still don't see
the "SeqLab" folder and Sequencing Browser icon please contact
juri.ahokas(ät)ktl.fi.
Review the raw data of your chromatograms
by SequenceScanner,
Sequencing
Analysis or with FinchTV software. The raw data view tells you a
lot about what's wrong in the sample:
- no signal: no template, no BigDye or no primer
- duplicate sequence peaks: failed PCR purification or
unspecific PCR
- fading signal: too much template DNA or too
much salt.
- Shadow peak for each base at position -1:
primer synthesis
- More to come soon -
BigDye
v3.1 chemistry allows dilution of original reactions of the protocol with
dilution buffer. Signal intensities may decrease but e.g. ABI3730xl
is able to produce high quality sequences with high dilution
factors. Motivation for diluting reagents is
decrease in
sequencing costs.
Dilution factor
used is dependent on template type and length. The
shorter the fragment the less BigDye 3.1 reagent
is needed in reaction. Excess use of BigDye3.1 reagent may
lead to dye blobs in chromatograms because sequencing reaction cleanup is
unable to remove all excess dye terminators from samples.
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