ESTP primers from Brown et al. (2001)

PCR primers were designed according to Harry et al. (1998) and Temesgen et al.(2000). Briefly, all primer sets were chosen to operate under a single thermal cycling regime which included a hot start and touchdown stages. In many cases, the “melting profile” of the amplification product predicted by the EST sequence was observed using MacMelt software (BioRad). This step allowed the effects of both primer position and GC clamping on melting profile to be examined and was used to optimize amplfication products for denaturing gradient gel electrophoresis.

Sequences of loblolly pine primer pairs from xylem ESTs suitable as anchor loci for comparative mapping.

FROM:
Brown, G.R, Kadel, E.E. III, Bassoni, D.L., Kiehne, K.L., Temesgen, B., van Buijtenen, J.P.,
Sewell, M.M., Marshall, K.A. and Neale, D.B.,  2001. Anchored reference loci in loblolly pine
(Pinus taeda L.) for integrating pine genomics. Genetics, 159:799-809.

^a  Clone refers to the loblolly pine xylem cDNA from which the EST sequence was derived. Primer sequences are given 5′ to 3′. Lower case letters denoted nucleotides added as GC-clamps and are not homologous to the corresponding EST sequence.
^b  GenBank Accession number.