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� 9 a rr,■•r biolinsi hts Bio-Trap - DNA Sampling Protocol SAMPLING INSTRUCTIONS Storage: It is important to minimize the amount of time that Bio-Trap Samplers are stored prior to being installed in the field.The physical properties of the Bio-Trap Samplers that make them an ideal medium for collecting microbes also increase the chances of microbial or chemical contamination.Bio-Trap Samplers need to remain sealed and refrigerated(not frozen)until they can be installed in the field. Note:Clean latex gloves(or similar)should be used at all times when handling Bio-Trap Samplers. Installation: • Prior to installing the Bio-Trap Sampler,the monitoring well may need to be purged if it has not been sampled in a while.If purging is necessary,MI recommends that three well volumes be removed to ensure contact with formation water and reduce well bore effect. • Attach the Bio-Trap Sampler's nylon loop(provided)to a nylon line(not provided)and suspend the Bio-Trap Sampler at a depth where significant contaminant concentrations exist.If no data is available on the vertical distribution of contaminants,then suspend the Bio-Trap Sampler in the middle of the saturated screened interval. • If large fluctuations in the water level are anticipated during the period of incubation,the Bio-Trap Sampler should be suspended from a float(contact MI for further details). Be sure not to suspend the Bio-Trap in the NAPL zone. • Once installed,incubation times can vary depending upon the scope of the project(routine monitoring and stable isotope probing(SIP)-3o days and"baited"- 6o days). Retrieval: • Open the monitoring well and pull up the Bio-Trap Sampler.Cut and remove the braided nylon line used to suspend the Bio-Trap Sampler. • Transfer the recovered Bio-Trap Sampler to labeled(well number and date)zippered bags,seal and then double bag in a larger(one-gallon)zippered bag, immediately place on blue ice in a cooler. • Repeat the above for all Bio-Trap Samplers from the site.Individual zippered bags containing the Bio-Trap Samplers can be placed in the same one-gallon zippered bag(if there is enough space). • A chain of custody(COC)form must be included with each shipment of samples. Hold time for this analysis is 24-48 hours. SHIPPING INSTRUCTIONS Packaging Samples: i. Samples should be shipped in a cooler with ice or blue ice for next day delivery.If regular ice is used,the ice should be double bagged. 2. A chain of custody form must be included with each shipment of samples. Access our chain of custody at www.microbe.com. Shipment for Weekday Delivery: Samples for weekday delivery should be shipped to: Sample Custodian Microbial Insights,Inc. 1o515 Research Drive Knoxville,TN 37932 (865)573-8188 Shipment for Saturday Delivery: Coolers to be delivered on Saturday must be sent to our FedEx Drop Location. To ensure proper handling the following steps must be taken: i. Fed Ex shipping label should be marked under(6)Special Handling,check Hold Saturday. 2. The cooler must be taped with FedEx SATURDAY tape. 3. The shipping label must be filled out with the Drop Location address below.Our laboratory name must be on the address label. 4.You MUST notify by email customerservice@microbe.com with the tracking number of the package on Friday(prior to 413m Eastern Time)to arrange for Saturday pickup. Please make sure you write"Saturday Delivery"in the subject line of the message.Without proper labeling and the tracking number,there is no guarantee that the samples will be collected. Samples for Saturday delivery should be shipped to: Microbial Insights,Inc. FedEx Drop Location io6oi Murdock Drive Knoxville,TN 37932 (865)3oo-8o53 10515 Research Drive Knoxville,TN 37932 Phone:865.573.8188 Fax:865.573.8133 www.microbe.com