Template Secondary Structure Promotes Polymerase Jumping During PCR Amplification
نویسندگان
چکیده
منابع مشابه
DNA damage promotes jumping between templates during enzymatic amplification.
Pairs of templates and primers were designed so that only recombination events would lead to amplification via the polymerase chain reaction. This approach reveals that lesions such as breaks, apurinic sites, and UV damage in a DNA template can cause the extending primer to jump to another template during the polymerase chain reaction. By comparing sequences of amplification products that were ...
متن کاملThe effect of template secondary structure on vaccinia DNA polymerase.
Vaccinia virus DNA polymerase will utilize a substrate consisting of phi X174 DNA primed with a strand of a unique restriction fragment, but the reaction is inefficient. Examination of the reaction products by alkaline agarose gel electrophoresis revealed a few discrete fragments, each corresponding to an extended primer strand. This result implies that specific barriers exist on the phi X174 t...
متن کاملErrors induced during PCR amplification
The Polymerase Chain Reaction (PCR) is one of the most widely used techniques in modern molecular biology to amplify a single or few copies of a piece of DNA sequences. The PCR products are used as inputs of many other applications such as diagnosis of diseases. Therefore the accuracy of the further analysis depends on the quality of the PCR outputs. In order to study the amplification process ...
متن کاملTemplate secondary structure can increase the error frequency of the DNA polymerase from Thermus aquaticus.
Amplification of portions of the intergenic spacer between the katE gene and cryptic cel operon of Escherichia coli was accomplished by the polymerase chain reaction using the DNA polymerase from Thermus aquaticus. Nine different segments were amplified and cloned without error, but one 83-bp fragment was amplified with a high error rate such that 32 of 34 selected clones had three or more nucl...
متن کاملThermostable Mismatch-Recognizing Protein MutS Suppresses Nonspecific Amplification during Polymerase Chain Reaction (PCR)
Polymerase chain reaction (PCR)-related technologies are hampered mainly by two types of error: nonspecific amplification and DNA polymerase-generated mutations. Here, we report that both errors can be suppressed by the addition of a DNA mismatch-recognizing protein, MutS, from a thermophilic bacterium. Although it had been expected that MutS has a potential to suppress polymerase-generated mut...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
ژورنال
عنوان ژورنال: BioTechniques
سال: 1999
ISSN: 0736-6205,1940-9818
DOI: 10.2144/99273st04