TempliPhi 100 扩增试剂盒

DNA from TempliPhi reactions consists of linear concatamers of the amplified plasmid sequence. Direct transformation of DNA into yeast may yield some transformants; results in bacterial are less efficient. To achieve optimal results with either organism, we recommend performing a restriction digest of the amplified DNA followed by ligation back into circular DNA.  

A TempliPhi could be used for amplification and enrichment of circular genomes, because it preferentially amplifies circular  DNA.

Yes. TempliPhi is designed to yield the same quantity of DNA regardless of the input quantity. Therefore, quantification is not necessary, and an aliquot can be added directly to sequencing reactions.

Phi29 polymerase extends from random priming sites and proceeds around the plasmid or circular DNA, displacing synthesizing strands in its path. Each generated strand can be in excess of 100 kb and will contain multiple side-by-side copies of the sequence in a long linear form (i.e., linear concatamers). Each amplified strand acts as new template and is further amplified.

TempliPhi has been used to amplify mitochondrial DNA. Rolling circle amplification does enrich for small circular DNA in terms of copy number over surrounding genomic DNA, but this will also be amplified to an extent.

Bacterial culture, colonies, or purified DNA can be used. TempliPhi can amplify picogram inputs, so avoid using too much input material because this may inhibit amplification.

TempliPhi can be used directly in most PCR reactions. However, purification and accurate quantitation may be preferred prior to sensitive applications.