title: Slowing PCR ramp speed reduces chimera formation from environmental samples.
abstract: Chimeric sequences falsely increase the apparent diversity within samples. To examine chimera formation in PCR products from environmental DNA, clone libraries were prepared using different ramp speeds to reach the designated temperatures for each step of the PCR program. Slowing the thermocycler ramp speed to 1 °C s(-1) reduced chimera formation.
authors.affiliation: Department of Biological Sciences, Campus Box 870344, University of Alabama, Tuscaloosa, AL 35487, United States.
authors.forename: Julia L
authors.initials: JL
authors.lastname: Stevens
authors.forename: Ronneshia L
authors.initials: RL
authors.lastname: Jackson
authors.forename: Julie B
authors.initials: JB
authors.lastname: Olson
mesh_terms: D016477:Artifacts; D004783:Environmental Microbiology; D008967:Molecular Biology / Q000379:methods*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA; D013048:Specimen Handling / Q000379:methods*; D013696:Temperature; D013997:Time Factors