A total of 158 municipality-supplied drinking water samples (20 L volume) were collected from different residential area of Delhi between periods of April, 2007 and May, 2010 in 25 L polypropylene container and carried to laboratory in minimal period of time. The samples were filtered and concentrated through 0.22 μm polyethersulfone pellicon-2 minifilter (Millipore Corporation, Billerica, MA01821) in a masterflex pellicon-2 miniholder using a Millipore vacuum pressure pump at 115V and 3 ψ. The filtration and concentration unit was rinsed with double-distilled water followed by 0.1N NaOH before and after each use to prevent cross contamination between two samples. Finally, 20–25 ml of sample was further centrifuged at 12,000 rpm for 20 min to concentrate it up to 0.5 ml volume.
Vacuum pressure pump
Manufactured by Merck Group
The Vacuum pressure pump is a laboratory equipment designed to create and maintain a vacuum. It generates a reduced air pressure environment within a confined space, enabling various applications that require controlled atmospheric conditions.
Automatically generated - may contain errors
2 protocols using vacuum pressure pump
1
Drinking Water Sampling and Concentration
2
Environmental Microbiome Profiling via Filtration
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Each water sample was aseptically filtered through a polyethersulfone membrane (0.22 μm pore size, 50 mm diameter) using vacuum-driven filters (Biofil) and a vacuum-pressure pump (Millipore). Filter membranes were cut into small pieces using a stainless steel scalpel and placed in sterile tubes. A control using ultrapure water was carried out to verify whether the membrane or filtration process could introduce any contamination. Bacterial genomic DNA was extracted from bacterial cells retained on each filter membrane using the DNeasy PowerSoil Kit (Qiagen), following the manufacturer’s methods. A reagent blank extraction was the control of the DNA extraction process. DNA sample concentration was measured using a Qubit fluorescence assay (Invitrogen). All DNA samples were concentrated in a vacufuge concentrator (Eppendorf) and sent for amplicon sequencing (16S rRNA, V4 region primers) on an Illumina HiSeq PE250 instrument at Novogene Bioinformatics Technology Co., Ltd. (Beijing, China). Since the controls, both from the filtration process and the DNA extraction resulted in negative PCR amplification, they were not further processed and were not sequenced (Additional file 2 ).
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