Anuran cross-species molecular sensors

摘要

Described herein are DNA primer sequences designed for the determination of gene or transcript information from Anuran species, and which may be used in studies for developmental and/or toxicity testing and for environmental toxicology or ecological assessment. Also described herein is a rapid, sensitive, high-throughput assay useful for supporting potential risk assessment across vertebrate clades, and that is also useful for evaluation of complex contaminant mixtures.

主权项

1. A frog tadpole metamorphosis assay method comprising a first phase and a second phase, wherein:
the first phase comprises exposing a tadpole to a compound of interest, removing first and second portions of the tadpole's tail, subjecting the first portion to an apoptosis assay, and the second phase comprises subjecting the second portion to an assay to detect shifts in a quantity of pre-determined mRNA transcripts, wherein the second phase further comprises subjecting the second portion to gPCR using a primer pair comprising a first primer and a second primer, wherein the primer pair is selected from the group consisting of primer pairs B, C, L, M:SEQSEQPrimerIDIDPairFirst PrimerNO:Second PrimerNO:BTACAGCTTCACCACCACAG 3TCCACATCTGCTGG 4AAGGTCTTTGCYTGGCGKCACTTTT  5ARCRGCACTGCGYC 6TGTALCAGGGGACAGAGAAAAGG23TGAGCTTTCTTGCC24TGACAGMGCTGCTGGTGTTGGTGART 25AGCATGTTGTCACC 26.RTTCC