It is recommended to check my sequence of interest for insertion sites on
the TargeTron Test Site before purchasing a kit. Why?
The test site will help you determine the number of potential insertion sites in
your sequence of interest. This is especially important for small DNA sequences
(400bp or less) as smaller sequences may not have potential insertion sites. The
locations of the insertion sites and the primer designs for those insertion sites
are only available upon purchase of the kit.
Is the cost of the primers included in the price of the TargeTron
kit?
No, the primers must be purchased separately. Primers can be easily purchased
through the website by using the link to Sigma-Genosys at the bottom of the
TargeTron Design Site output page.
When designing my primers to mutate the intron using the TargeTron Design
Site, the site suggests choosing the primer sets with the higher S-values and the
lower E-values. What does this mean?
DNA target sequences were scored (S-value) using a
log-odds score, which measures whether the probability that the sequence
generated by the model is greater than the probability that the sequence was
generated by chance (null model). A positive score means that the model fits the
group II intron target sequence better than chance (null model). Therefore, the
higher the score (S-value), the better the chance
that the target site is "real" and that the intron will insert efficiently. (Perutka et. al. 2003. Use of Computer-designed Group II Introns to Disrupt Escherichia Coli DexH/D-box Protein and DNA Helicase Genes. J. Mol. Bio.336, 421-429).
The E-value estimates the expected number of false
positives at or above the score (S-value).
Can I save the primer sequences generated by the TargeTron Design
Site?
Yes. There is a link to download the primer sequences to a text file for future
reference or to order primers. Downloading your results is important. Once you
leave the TargeTron Design Site, you will not be able to access the primer
sequences again without using another access.
Look for the text below and the link to download results to a text file.
Below are the potential insertion sites and the primer sets corresponding to the design options. Please download the results for reference and/or to order primers. Once this window is closed, the results cannot be viewed without using an additional access.
