Eliminating primers from completed polymerase chain reactions with exonuclease VII.
نویسندگان
چکیده
Single-stranded oligonucleotide primers can be efficiently removed after PCR using E.coli exonuclease VII. Even only a few molecules of double stranded PCR product are unaffected by a treatment which eliminates 20 picomoles of primer in the presence of 500 ng of denatured genomic DNA. Exonuclease VII treatment is rapid and could simplify complicated multistep PCR protocols.
منابع مشابه
Taq DNA polymerase extension of internal primers blocks polymerase chain reactions allowing differential amplification of molecules with identical 5' and 3' ends.
Polymerase chain reaction (PCR) methodology (1) has become a routine method for selectively amplifying segments of DNA from a wide variety of sources. Amplification of specific sequences is dependent upon an exact match between the template DNA and the oligonucleotide primers. Mismatches at the 3' terminus lead to greatly reduced amplification, with no detectable product when amplified under th...
متن کاملThe use of exonuclease III for polymerase chain reaction sterilization.
The carryover of previously amplified sequences (amplicons) into new PCR reactions is a serious problem. Recently Furrer et al. reported a 'pre-PCR' sterilization technique using DNase I or restriction enzymes for contamination control (1). Unfortunately, these methods require the reaction tube to be re-opened to add target DNA and Taq polymerase following the enzymatic treatment (providing an ...
متن کاملDNA polymerase fidelity and the polymerase chain reaction.
High-fidelity DNA synthesis conditions are those that exploit the inherent ability of polymerases to discriminate against errors. This review has described several experimental approaches for controlling the fidelity of enzymatic DNA amplification. One of the most important parameters to consider is the choice of which polymerase to use in PCR. As demonstrated by the data in Tables 2 and 3, hig...
متن کاملDeconstructing the Polymerase Chain Reaction: Understanding and Correcting Bias Associated with Primer Degeneracies and Primer-Template Mismatches
The polymerase chain reaction (PCR) is sensitive to mismatches between primer and template, and mismatches can lead to inefficient amplification of targeted regions of DNA template. In PCRs in which a degenerate primer pool is employed, each primer can behave differently. Therefore, inefficiencies due to different primer melting temperatures within a degenerate primer pool, in addition to misma...
متن کاملOligoribonucleotide (ORN) Interference-PCR (ORNi-PCR): A Simple Method for Suppressing PCR Amplification of Specific DNA Sequences Using ORNs
Polymerase chain reaction (PCR) amplification of multiple templates using common primers is used in a wide variety of molecular biological techniques. However, abundant templates sometimes obscure the amplification of minor species containing the same primer sequences. To overcome this challenge, we used oligoribonucleotides (ORNs) to inhibit amplification of undesired template sequences withou...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Nucleic acids research
دوره 19 11 شماره
صفحات -
تاریخ انتشار 1991