Hello EvolDir members, I sent the email below in October in the hopes of finding the complete reference to the described paper. I thank everybody who took the time to respond. I have not had any success in finding the reference, though I've had interest from others who would also like the reference to the described paper. I was also emailed suggested reading material, which is listed below. The technique in the Brownstein et al. (1996) paper is useful to remove stutter caused by + A artifacts, though I believe the stutter in my case is not caused by this. I found the last paper listed particularly useful as a guide in reducing stutter thought to be caused by strand slippage, though the described novel technique of reducing denaturation temperature was unsuccessful in the few dinucleotide loci I attempted. I'd be interested to hear if anybody has had success (or no success) with using a reduced denaturation temperature to decrease stutter, particularly for dinucleotides as the paper only tested trinucleotides. Brownstein, M., Carpten, J., Smith, J. (1996) Modulation of non-templated nucleotide addition by Taq DNA polymerase: primer modifications that facilitate genotyping. Biotechniques 20: 1004-1010 Munoz-Fuentes, V., Gyllenstrand, N., Negro, J., Green, A., Vila, C. (2005) Microsatellite markers for two stifftail ducks: the white-headed duck, Oxyura leucocephala, and the ruddy duck, O. jamaicensis. Molecular Ecology Notes 5:263-265 (which uses the technique described in the above paper) Krzyzosiak, W., Olejniczak, M. (2006) Genotyping of simple sequence repeats - factors implicated in shadow band generation revisited. Electrophoresis 27: 3724-3734 Kind regards, Catherine Hello EvolDir members, I'm trying to find (with no luck) the original Biotechniques paper which describes the technique of using greatly abbreviated PCR times (e.g. 2/2/8 s) at each step to decrease microsatellite stutter. This technique was described in the response of Andrew DeWoody in 2006 (see below for quoted response) to an EvolDir member that asked about how to decrease stutter. I do not know the complete reference of this paper, and as Andrew read the paper a while ago he is also unsure of the reference though he thinks the paper might be about hymenopterans. Does anybody know the full reference of this paper or (preferentially) can they email the paper as an attachment to catherine.attard@students.mq.edu.au? Many thanks and kind regards, Catherine EvolDir response: "You might try greatly abbreviated times at each step in PCR (e.g., 2 sec denaturation, 2 sec annealing, 8 sec extension). The original paper describing this was published in Biotechniques about 6 or 7 years ago; see also the attached reprint. [We had cited the Biotechniques paper in the bass reprint, but the citation was ultimately cut due to strict page limits in PRS.] In my experience, the approach either works very well or fails completely (on a locus by locus basis). If you get amplifications, the stutter is usually greatly reduced. DeWoody et al. (2000) Proc. R. Soc. Lond. B 267:2431-2437." catherine.attard@students.mq.edu.au