http://lod.bco-dmo.org/id/dataset/553477
eng; USA
utf8
dataset
Highest level of data collection, from a common set of sensors or instrumentation, usually within the same research project
Biological and Chemical Oceanography Data Management Office (BCO-DMO)
Unavailable
508-289-2009
WHOI MS#36
Woods Hole
MA
02543
USA
info@bco-dmo.org
http://www.bco-dmo.org
Monday - Friday 8:00am - 5:00pm
For questions regarding this resource, please contact BCO-DMO via the email address provided.
pointOfContact
2015-03-13
ISO 19115-2 Geographic Information - Metadata - Part 2: Extensions for Imagery and Gridded Data
ISO 19115-2:2009(E)
T-RFLP, response of the microbial community to coral spawning, lagoon and reef flats in Kaneohe Bay, Oahu, Hawaii during 2006 - 2007 (MiCoDe project)
2015-03-12
publication
2015-03-12
revision
BCO-DMO Linked Data URI
2015-03-12
creation
http://lod.bco-dmo.org/id/dataset/553477
Michael S. Rappé
University of Hawaiʻi at Mānoa
principalInvestigator
Biological and Chemical Oceanography Data Management Office (BCO-DMO)
Unavailable
508-289-2009
WHOI MS#36
Woods Hole
MA
02543
USA
info@bco-dmo.org
http://www.bco-dmo.org
Monday - Friday 8:00am - 5:00pm
For questions regarding this resource, please contact BCO-DMO via the email address provided.
publisher
Cite this dataset as: Rappé, M. S. (2015) T-RFLP, response of the microbial community to coral spawning, lagoon and reef flats in Kaneohe Bay, Oahu, Hawaii during 2006 - 2007 (MiCoDe project). Biological and Chemical Oceanography Data Management Office (BCO-DMO). (Version 2015-03-12) Version Date 2015-03-12 [if applicable, indicate subset used]. http://lod.bco-dmo.org/id/dataset/553477 [access date]
T-RFLP, response of the microbial community to coral spawning, lagoon and reef flats, Oahu, 2006-7 Dataset Description: <p>The response of the microbial community to coral spawning was investigated over a period of 18 mo, from January 2006 to July 2007, in reef flat and lagoon environments of a subtropical embayment (Kaneohe Bay, Oahu, Hawaii, USA). The composition of the bacterioplankton community was characterized using terminal restriction fragment length polymorphism (T-RFLP) analysis of bacterial small-subunit (SSU) ribosomal RNA genes in parallel with measurements of microbial cell abundances, bacterial production via 3H-leucine incorporation, and seawater biochemical parameters. Abundance of coral eggs, zooplankton, and bacterial, and chlorophyll&nbsp;were measured along with&nbsp;nutrient analysis and&nbsp;physical parameters.</p>
<p><strong>Related Reference:</strong></p>
<p>These data are published in&nbsp;Apprill, A. M., and M. S. Rappé (2011) Response of the microbial community to coral spawning in lagoon and reef flat environments of Hawaii, USA. Aquatic Microbial Ecology, 62:251-266.</p> Methods and Sampling: <p><strong>T-RFLP analysis of bacterial SSU rRNA genes.</strong> Approximately 1 l of seawater was filtered through a GF/A glass microfiber membrane pre-filter (1.6 µm nominal pore size, Whatman International) followed by a 13 mm diameter, 0.2 µm pore-sized polyethersulfone membrane (Supor 200, Pall Gelman). Filters were stored at -80°C in DNA lysis buffer (20 mM Tris-HCl pH 8.0, 2 mM EDTA pH 8.0, 1.2% v/v Triton X100) (Suzuki et al. 2001). Genomic DNA was extracted from the 0.2 µm pore-sized polyethersulfone membranes using a modified version of the DNeasy Tissue kit (Qiagen) (Becker et al. 2007), and quantified using a PicoGreen fluorescent assay (Invitrogen) on a SpectraMax M2 plate reader (Molecular Devices). For terminal restriction fragment length polymorphism (T-RFLP) analysis (Liu et al. 1997), bacterioplankton small-subunit (SSU) ribosomal RNA (rRNA) genes (including those of heterotrophic bacteria, cyanobacteria, and eukaryotic plastids) were first amplified via the polymerase chain reaction (PCR) using the bacterial primers 27F-B-FAM (5’-AGRGTTYGATYM TGGCTCAG-3’) and 519R (5’-GWATTACCGCGGCKGCTG- 3’), with ‘FAM’ indicating 5’ end-labeling of the 27F-B primer with the 6-carboxyfluorescein (FAM) fluorochrome. Each 50 µl PCR reaction contained 0.625 U of PicoMaxx high-fidelity DNA polymerase (Stratagene), 1× PicoMaxx reaction buffer, 200 µM of each deoxynucleoside triphosphate (dNTP), 200 nM of each primer and 10 ng of environmental genomic DNA template. After an initial denaturation step at 95°C for 5 min, the reaction conditions were: 24 cycles of 95°C denaturation for 30 s, 55°C annealing for 1 min, and 72°C extension for 2 min, concluding with an extension at 72°C for 20 min. The reactions were performed in a MyCycler Personal Thermal Cycler (Bio-Rad Laboratories). Amplification products were purified using the QIAquick PCR Purification Kit (Qiagen), and subsequently restricted in a 10 µl reaction containing 100 ng of purified amplification product, 2 µg of bovine serum albumin (BSA), 1× enzymatic reaction buffer, and 5 units of HaeIII restriction endonuclease (Promega) for 7 h at 37°C. Restriction digests were purified using the QIAquick Nucleotide Removal Kit (Qiagen), and 30 ng µl-1 of each product was subsequently electrophoresed on an ABI 3100 Genetic Analyzer (Applied Biosystems). GeneMapper software (Applied Biosystems) was used to estimate the length (in base pairs) and relative abundance of the resulting fragments. Operational taxonomic units (OTUs) were identified as terminal restriction fragments (T-RFs) detected between 33 and 550 bp in length. To account for small differences in the amount of DNA loaded on the ABI 3100, data were normalized by excluding peaks that contributed to &lt;0.05% of the total peak area for each sample (Sait et al. 2003). An average of 2 peaks were removed from the samples, with a minimum of no peaks and a maximum of 19 peaks removed. An in-house dataset linking cloned bacterial and plastid SSU rRNA gene sequences to T-RFLP profiles from Kaneohe Bay seawater was analyzed in order to putatively identify T-RFs of interest (Yeo et al. unpubl.).</p>
Funding provided by NSF Division of Ocean Sciences (NSF OCE) Award Number: OCE-0928806 Award URL: https://www.nsf.gov/awardsearch/show-award?AWD_ID=0928806
completed
Michael S. Rappé
University of Hawaiʻi at Mānoa
808-372-2394
University of Hawaii at Manoa - Hawaii Institute of Marine Biology 46-007 Lilipuna Rd
Kaneohe
HI
96744
USA
rappe@hawaii.edu
pointOfContact
asNeeded
Dataset Version: 2015-03-12
Unknown
sample_id
date_coll
time_coll
site
location
lat
lon
depth
production
egg_abund
zoop_biomass
TOC
PO4
NO3_NO2
SiO4
NO2
NH4
chl_a
Synech_abund
picoeuk_abund
nonpig_bact_abund
temp
sal
O2
pH
turbidity
tide
TRF_length
abund
plate reader
Thermal Cycler
theme
None, User defined
sample identification
date
time of day
site
site description
latitude
longitude
depth
production
abundance
dry_wgt_globec
total organic Carbon
reactive phosphorus (PO4)
nitrate plus nitrite
Silicate (reactive silica)
Nitrite
Ammonium
chlorophyll a
water temperature at measurement depth
salinity
dissolved Oxygen
pH
turbidity
tide
No BCO-DMO term
featureType
BCO-DMO Standard Parameters
Automated DNA Sequencer
plate reader
Thermal Cycler
instrument
BCO-DMO Standard Instruments
Rappe_2006
service
Deployment Activity
Kaneohe Bay, Oahu, Hawaii, USA
place
Locations
otherRestrictions
otherRestrictions
Access Constraints: none. Use Constraints: Please follow guidelines at: http://www.bco-dmo.org/terms-use Distribution liability: Under no circumstances shall BCO-DMO be liable for any direct, incidental, special, consequential, indirect, or punitive damages that result from the use of, or the inability to use, the materials in this data submission. If you are dissatisfied with any materials in this data submission your sole and exclusive remedy is to discontinue use.
The Development of Microbial Associations in Major Reef Building Corals of the Pacific Ocean
http://www.soest.hawaii.edu/oceanography/faculty/rappe/RappeLab/CAM.html
The Development of Microbial Associations in Major Reef Building Corals of the Pacific Ocean
<p><em>Description from NSF award abstract:</em><br />
Fundamental to the study of coral-microbial associations is an understanding of when and how the relationships are established, and their specificity. Recently, the investigators provided the first evidence of a specific association between an early life history stage of a coral (<em>Pocillopora meandrina</em>) and a particular group of bacteria (Jannaschia lineage of the Roseobacter clade of Alphaproteobacteria). They will be expanding this work by examining the onset of microbial associations in key reef building corals from Hawaii in the North Pacific Ocean and Moorea, French Polynesia, in the South Pacific Ocean. Understanding the onset, specificity and function of the microbial community associated with these coral species is necessary to understand and predict the coral holobiont response to a changing environment.</p>
<p>The main objectives of this proposal are to:<br />
1. Use cultivation independent techniques to identify and quantify microorganisms associated with several major reef building corals of Hawaii in the North Pacific Ocean that represent a variety of reproductive strategies (brooding and broadcast spawning) and differing modes of zooxanthellae symbiont transmission (vertical vs. horizontal), throughout the reproductive cycle, early developmental stages, and post-settlement stages of each.<br />
2. Collect and analyze similar samples from the same (or similar) species of coral found in the South Pacific Ocean in Moorea, French Polynesia, in order to assess whether the associations documented in objective 1 are localized to Hawaii, or broadly distributed across the Pacific and likely to represent common features of coral development.<br />
3. Use fluorescence in situ hybridization to enumerate cells of the Jannaschia lineage of the Roseobacter clade throughout the development cycle of P. meandrina collected in Hawaii. The PIs will expand this objective to include other coral species, target bacteria, and/or geographic location as they identify additional associations.<br />
4. Isolate microorganisms prevalent in cultivation-independent surveys of P. meandrina-associated microbial communities (e.g. Jannaschia sp.) by the application of novel culturing techniques, in order to develop model systems for the investigation of coral-microbe interactions.</p>
<p>Coral reefs are in decline as a result of increasing environmental stress due to anthropogenic activity, and there is now considerable evidence indicating that they are under threat from the effects of rising sea surface temperature and ocean acidification. Microorganisms associated with corals are thought to play a variety of potentially important roles in maintaining the health and resiliency of the coral host, and advances in methodology primarily driven by developments in the field of molecular biology are facilitating growing insight into this association. Much of coral-microbial research is focused on the contribution of microorganisms to disease and bleaching, and is focused almost exclusively on adult coral colonies. This study will provide unique information on the manner in which microorganisms interact with healthy corals throughout their developmental cycle, the specificity of these relationships, how they are initiated, and their distribution and frequency in nature.</p>
MiCoDe
largerWorkCitation
project
eng; USA
biota
oceans
Kaneohe Bay, Oahu, Hawaii, USA
2015-03-12
Kaneohe Bay, HI, USA; and Cook’s Bay, Moorea, French Polynesia
0
BCO-DMO catalogue of parameters from T-RFLP, response of the microbial community to coral spawning, lagoon and reef flats in Kaneohe Bay, Oahu, Hawaii during 2006 - 2007 (MiCoDe project)
Biological and Chemical Oceanography Data Management Office (BCO-DMO)
Unavailable
508-289-2009
WHOI MS#36
Woods Hole
MA
02543
USA
info@bco-dmo.org
http://www.bco-dmo.org
Monday - Friday 8:00am - 5:00pm
For questions regarding this resource, please contact BCO-DMO via the email address provided.
pointOfContact
http://lod.bco-dmo.org/id/dataset-parameter/553498.rdf
Name: sample_id
Units: unitless
Description: <p>sample identification</p>
http://lod.bco-dmo.org/id/dataset-parameter/553499.rdf
Name: date_coll
Units: yyyy-mm-dd
Description: <p>date collected</p>
http://lod.bco-dmo.org/id/dataset-parameter/553500.rdf
Name: time_coll
Units: HH:MM
Description: <p>time collected</p>
http://lod.bco-dmo.org/id/dataset-parameter/553501.rdf
Name: site
Units: unitless
Description: <p>site</p>
http://lod.bco-dmo.org/id/dataset-parameter/553502.rdf
Name: location
Units: unitless
Description: <p>locations of the sites</p>
http://lod.bco-dmo.org/id/dataset-parameter/553503.rdf
Name: lat
Units: decimal degrees
Description: <p>latitude; north is positive</p>
http://lod.bco-dmo.org/id/dataset-parameter/553504.rdf
Name: lon
Units: decimal degrees
Description: <p>longitude; east is positive</p>
http://lod.bco-dmo.org/id/dataset-parameter/553505.rdf
Name: depth
Units: meters
Description: <p>depth</p>
http://lod.bco-dmo.org/id/dataset-parameter/553506.rdf
Name: production
Units: ug C L-1 hr-1
Description: <p>carbon production</p>
http://lod.bco-dmo.org/id/dataset-parameter/553507.rdf
Name: egg_abund
Units: eggs/meter^3
Description: <p>coral egg abundance</p>
http://lod.bco-dmo.org/id/dataset-parameter/553508.rdf
Name: zoop_biomass
Units: mg/m3
Description: <p>zooplankton dry weight</p>
http://lod.bco-dmo.org/id/dataset-parameter/553509.rdf
Name: TOC
Units: uM
Description: <p>total organic carbon</p>
http://lod.bco-dmo.org/id/dataset-parameter/553510.rdf
Name: PO4
Units: uM
Description: <p>phosphate</p>
http://lod.bco-dmo.org/id/dataset-parameter/553511.rdf
Name: NO3_NO2
Units: uM
Description: <p>nitrate and nitrite</p>
http://lod.bco-dmo.org/id/dataset-parameter/553512.rdf
Name: SiO4
Units: uM
Description: <p>silicate</p>
http://lod.bco-dmo.org/id/dataset-parameter/553513.rdf
Name: NO2
Units: uM
Description: <p>nitrite</p>
http://lod.bco-dmo.org/id/dataset-parameter/553514.rdf
Name: NH4
Units: uM
Description: <p>ammonium</p>
http://lod.bco-dmo.org/id/dataset-parameter/553515.rdf
Name: chl_a
Units: (ug/L)
Description: <p>chlorophyll a</p>
http://lod.bco-dmo.org/id/dataset-parameter/553516.rdf
Name: Synech_abund
Units: cells/ml
Description: <p>Synechococcus abundance</p>
http://lod.bco-dmo.org/id/dataset-parameter/553517.rdf
Name: picoeuk_abund
Units: cells/ml
Description: <p>picoeukaryote abundance</p>
http://lod.bco-dmo.org/id/dataset-parameter/553518.rdf
Name: nonpig_bact_abund
Units: cells/ml
Description: <p>non-pigmented bacterioplankton</p>
http://lod.bco-dmo.org/id/dataset-parameter/553519.rdf
Name: temp
Units: degrees Celsius
Description: <p>temperature</p>
http://lod.bco-dmo.org/id/dataset-parameter/553520.rdf
Name: sal
Units: ppt
Description: <p>salinity</p>
http://lod.bco-dmo.org/id/dataset-parameter/553521.rdf
Name: O2
Units: mg/L
Description: <p>dissolved oxygen</p>
http://lod.bco-dmo.org/id/dataset-parameter/553522.rdf
Name: pH
Units: unitless
Description: <p>pH</p>
http://lod.bco-dmo.org/id/dataset-parameter/553523.rdf
Name: turbidity
Units: turbidity units
Description: <p>turbidity</p>
http://lod.bco-dmo.org/id/dataset-parameter/553524.rdf
Name: tide
Units: meters above mean low water
Description: <p>tide height</p>
http://lod.bco-dmo.org/id/dataset-parameter/553579.rdf
Name: TRF_length
Units: base pairs
Description: <p>terminal restriction fragment length</p>
http://lod.bco-dmo.org/id/dataset-parameter/553580.rdf
Name: abund
Units: TRF's
Description: <p>abundance of the TRF</p>
GB/NERC/BODC > British Oceanographic Data Centre, Natural Environment Research Council, United Kingdom
Biological and Chemical Oceanography Data Management Office (BCO-DMO)
Unavailable
508-289-2009
WHOI MS#36
Woods Hole
MA
02543
USA
info@bco-dmo.org
http://www.bco-dmo.org
Monday - Friday 8:00am - 5:00pm
For questions regarding this resource, please contact BCO-DMO via the email address provided.
pointOfContact
5670574
https://datadocs.bco-dmo.org/file/KAAGoPwSlk7WK/spawn_2011_r2col.csv
spawn_2011_r2col.csv
Primary data file for dataset ID 553477
download
https://osprey.bco-dmo.org/dataset/553477/data/download
download
onLine
dataset
<p><strong>T-RFLP analysis of bacterial SSU rRNA genes.</strong> Approximately 1 l of seawater was filtered through a GF/A glass microfiber membrane pre-filter (1.6 µm nominal pore size, Whatman International) followed by a 13 mm diameter, 0.2 µm pore-sized polyethersulfone membrane (Supor 200, Pall Gelman). Filters were stored at -80°C in DNA lysis buffer (20 mM Tris-HCl pH 8.0, 2 mM EDTA pH 8.0, 1.2% v/v Triton X100) (Suzuki et al. 2001). Genomic DNA was extracted from the 0.2 µm pore-sized polyethersulfone membranes using a modified version of the DNeasy Tissue kit (Qiagen) (Becker et al. 2007), and quantified using a PicoGreen fluorescent assay (Invitrogen) on a SpectraMax M2 plate reader (Molecular Devices). For terminal restriction fragment length polymorphism (T-RFLP) analysis (Liu et al. 1997), bacterioplankton small-subunit (SSU) ribosomal RNA (rRNA) genes (including those of heterotrophic bacteria, cyanobacteria, and eukaryotic plastids) were first amplified via the polymerase chain reaction (PCR) using the bacterial primers 27F-B-FAM (5’-AGRGTTYGATYM TGGCTCAG-3’) and 519R (5’-GWATTACCGCGGCKGCTG- 3’), with ‘FAM’ indicating 5’ end-labeling of the 27F-B primer with the 6-carboxyfluorescein (FAM) fluorochrome. Each 50 µl PCR reaction contained 0.625 U of PicoMaxx high-fidelity DNA polymerase (Stratagene), 1× PicoMaxx reaction buffer, 200 µM of each deoxynucleoside triphosphate (dNTP), 200 nM of each primer and 10 ng of environmental genomic DNA template. After an initial denaturation step at 95°C for 5 min, the reaction conditions were: 24 cycles of 95°C denaturation for 30 s, 55°C annealing for 1 min, and 72°C extension for 2 min, concluding with an extension at 72°C for 20 min. The reactions were performed in a MyCycler Personal Thermal Cycler (Bio-Rad Laboratories). Amplification products were purified using the QIAquick PCR Purification Kit (Qiagen), and subsequently restricted in a 10 µl reaction containing 100 ng of purified amplification product, 2 µg of bovine serum albumin (BSA), 1× enzymatic reaction buffer, and 5 units of HaeIII restriction endonuclease (Promega) for 7 h at 37°C. Restriction digests were purified using the QIAquick Nucleotide Removal Kit (Qiagen), and 30 ng µl-1 of each product was subsequently electrophoresed on an ABI 3100 Genetic Analyzer (Applied Biosystems). GeneMapper software (Applied Biosystems) was used to estimate the length (in base pairs) and relative abundance of the resulting fragments. Operational taxonomic units (OTUs) were identified as terminal restriction fragments (T-RFs) detected between 33 and 550 bp in length. To account for small differences in the amount of DNA loaded on the ABI 3100, data were normalized by excluding peaks that contributed to &lt;0.05% of the total peak area for each sample (Sait et al. 2003). An average of 2 peaks were removed from the samples, with a minimum of no peaks and a maximum of 19 peaks removed. An in-house dataset linking cloned bacterial and plastid SSU rRNA gene sequences to T-RFLP profiles from Kaneohe Bay seawater was analyzed in order to putatively identify T-RFs of interest (Yeo et al. unpubl.).</p>
Specified by the Principal Investigator(s)
<p><strong>Statistical analyses.</strong> Normalized T-RFLP data were analyzed using PC-ORD software (MjM Software Design). Multi-response permutation procedure (MRPP) analysis was employed to test for significant differences in T-RFLP bacterioplankton communities between sites (McCune &amp; Grace 2002). Species indicator analysis was used to identify T-RFs which were significantly different between parameters (Dufrene &amp; Legendre 1997, McCune &amp; Grace 2002). Non-metric multidimensional scaling (NMS) analysis was utilized to explore relationships between individual T-RFLP samples, and was conducted using the Sorenson (Bray- Curtis) distance measure with slow and thorough autopilot criteria (McCune &amp; Grace 2002, Fierer &amp; Jackson 2006). Mantel tests using Sorenson (Bray-Curtis) distance measures and Monte Carlo simulations (1000 randomized runs) were employed to test the relationship between quantitative abiotic and biotic parameters against the T-RFLP profiles.</p>
<p><strong>BCO-DMO Processing:</strong></p>
<p>original file: Apprill_AME_2011_data_v2.xlsx<br />
- added conventional header with dataset name, PI name, source information<br />
- renamed parameters to BCO-DMO standard<br />
- replaced blank space with _, * with nd, and removed commas<br />
- sorted by site, date, sample_id<br />
- transposed trf rows to columns<br />
- combined metadata with trf data<br />
- ran rows-to-columns.pl script to transform T-RF length and abundance rows to columns</p>
Specified by the Principal Investigator(s)
asNeeded
7.x-1.1
Biological and Chemical Oceanography Data Management Office (BCO-DMO)
Unavailable
508-289-2009
WHOI MS#36
Woods Hole
MA
02543
USA
info@bco-dmo.org
http://www.bco-dmo.org
Monday - Friday 8:00am - 5:00pm
For questions regarding this resource, please contact BCO-DMO via the email address provided.
pointOfContact
PI Supplied Instrument Name: PI Supplied Instrument Description:ABI 3100 Genetic Analyzer (Applied Biosystems, Foster City, CA) Instrument Name: Automated DNA Sequencer Instrument Short Name:Automated Sequencer Instrument Description: A DNA sequencer is an instrument that determines the order of deoxynucleotides in deoxyribonucleic acid sequences.
plate reader
plate reader
PI Supplied Instrument Name: plate reader PI Supplied Instrument Description:SpectraMax M2 plate reader (Molecular Device Corp., Sunnyvale, CA, USA) Instrument Name: plate reader Instrument Short Name: Instrument Description: Plate readers (also known as microplate readers) are laboratory instruments designed to detect biological, chemical or physical events of samples in microtiter plates. They are widely used in research, drug discovery, bioassay validation, quality control and manufacturing processes in the pharmaceutical and biotechnological industry and academic organizations. Sample reactions can be assayed in 6-1536 well format microtiter plates. The most common microplate format used in academic research laboratories or clinical diagnostic laboratories is 96-well (8 by 12 matrix) with a typical reaction volume between 100 and 200 uL per well. Higher density microplates (384- or 1536-well microplates) are typically used for screening applications, when throughput (number of samples per day processed) and assay cost per sample become critical parameters, with a typical assay volume between 5 and 50 µL per well. Common detection modes for microplate assays are absorbance, fluorescence intensity, luminescence, time-resolved fluorescence, and fluorescence polarization. From: http://en.wikipedia.org/wiki/Plate_reader, 2014-09-0-23.
Thermal Cycler
Thermal Cycler
PI Supplied Instrument Name: Thermal Cycler PI Supplied Instrument Description:MyCycler thermal cycler (Bio-Rad Laboratories, Hercules, CA, USA) Instrument Name: Thermal Cycler Instrument Short Name:Thermal Cycler Instrument Description: A thermal cycler or "thermocycler" is a general term for a type of laboratory apparatus, commonly used for performing polymerase chain reaction (PCR), that is capable of repeatedly altering and maintaining specific temperatures for defined periods of time. The device has a thermal block with holes where tubes with the PCR reaction mixtures can be inserted. The cycler then raises and lowers the temperature of the block in discrete, pre-programmed steps. They can also be used to facilitate other temperature-sensitive reactions, including restriction enzyme digestion or rapid diagnostics.
(adapted from http://serc.carleton.edu/microbelife/research_methods/genomics/pcr.html)
Deployment: Rappe_2006
Rappe_2006
Hawaii_reef
shoreside
Rappe_2006
Michael S. Rappé
University of Hawaiʻi at Mānoa
Hawaii_reef
shoreside