General Considerations For Oligonucleotide Design
80-100 bp oligonucleotides can be generated as single molecules, or pairs of oligonucleotides that are completely overlapping or partially overlapping. The type of oligonucleotide recommended depends on the type of mutation and the distance from the CORE cassette integration site a mutation is desired.
Longer oligonucleotides lead to increased transformation efficiency. Fully complementary oligonucleotides pairs lead to 5-10 fold increase in efficiency compared to single oligonucleotides and are recommended for all applications. However, they provide a small window of mutagenesis of only 20-40 bases from the CORE cassette. To increase the window of mutagenesis, oligonucleotide pairs with a 20 bp overlap can be used, and these allow up to 100 bp upstream and downstream of the CORE integration site to be targeted. However, they transform approximately 6 times less efficiently. To increase the efficiency, partly overlapping oligonucleotides can be extended in vitro.
Read more about this topic: Delitto Perfetto
Famous quotes containing the words general and/or design:
“The happiest conversation is that of which nothing is distinctly remembered but a general effect of pleasing impression.”
—Samuel Johnson (1709–1784)
“We find that Good and Evil happen alike to all Men on this Side of the Grave; and as the principle Design of Tragedy is to raise Commiseration and Terror in the Minds of the Audience, we shall defeat this great End, if we always make Virtue and Innocence happy and successful.”
—Joseph Addison (1672–1719)