The Methods we used right now.
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The first primer for primer walking is the vector. Then if we want to
continue the primer walking, we have to select the next primer.
<p>
Right now, we are using several other software to finish selection octamer
primer from the octamer primer library. 
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<a href= dis.gif> Re2 project </a>
<p>
Sequence Methods:<p>
 Automsated sequencing reactions were performed using ABI BigDye
Terminator Cycle
Sequencing Ready Reaction kit with AnpliTaq DNA Polymerase. Sequencing
reactions were precipitated, pellets were resuspended by buffer, and was
loaded onto a sequencing gel. The data was collected by an ABI PRISM 377
DNA Sequencher.
<p>
 Using Sequencher 3.0 (GeneCodes, Inc.), the location of
ocatamers identified by this comparison was presented on the sequence,
using alignment settings that requirea 100% match and a 8 base overlap.
After the positions of the octmaer matches were identified, the
information was exported to a Microsoft Excel spreadsheet for further
analysis.
<p>
 After that, we have to analysis of template secondary structure.
The template DNA sequence was analyzed for potential hairpin structure
using OLIGO 5.0. After the location of the octamer was identified, a 40
base region of the template (centering on the ocatamer) was analyzed for
potential hairpin structures. 

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