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DATA COLLECTION CONSTRAINTS FOR THE USE OF LENGTH HETEROGENEITY POLYMERASE CHAIN REACTION (LH-PCR) AS AN INDICATOR OF STREAM SANITARY AND ECOLOGICAL CONDITION
Citation:
Bracken, C. L., K. G. Field, AND A. K. Harding. DATA COLLECTION CONSTRAINTS FOR THE USE OF LENGTH HETEROGENEITY POLYMERASE CHAIN REACTION (LH-PCR) AS AN INDICATOR OF STREAM SANITARY AND ECOLOGICAL CONDITION. Presented at regional Water Quality Conference, Vancouver, WA, February 20-21, 2001.
Description:
This study is part of a larger project for the development of bacterial indicators of stream sanitary and ecological condition. Here we report preliminary research on the use of Length Heterogeneity Polymerase Chain Reaction (LH-PCR), which discriminates among 16S rRNA genes based on differences in the length of PCR products in a mixed environmental sample. An automated DNA sequencer yields a community profile of 16S rRNA gene products known as an electropherogram. This technology offers a fairly quick and inexpensive look at in situ bacterial community diversity and has been found to vary along certain environmental gradients.
One of the limitations posed by the traditional use of coliform plate count data to assess fecal pollution is the need to analyze samples within a 24-hour period. However, it may be possible to use LH-PCR as an initial screening tool to assess the overall bacterial load of streams, offering a cost effective method of identifying areas of concern on a regional scale. This phase of research addresses issues concerning the minimum volume of water needed per sample, as well as the maximum holding time and temperature of filtered samples in Guanidine Thiocyanate prior to DNA extraction in the lab. This work will be used to develop the data collection protocol for a field study comparing traditional public health indicators, including fecal coliform plate counts, to LH-PCR electropherograms generated from samples collected in Willamette Valley streams.