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Sample GSM1139165 Query DataSets for GSM1139165
Status Public on Jun 20, 2013
Title Water column DNA_Arabian Sea_Station 1_175m
Sample type mixed
 
Channel 1
Source name Universal Internal Standard + 70-mer
Organism synthetic construct
Characteristics sample type: reference
Extracted molecule other
Extraction protocol DNA was extracted from the filters using the PureGene DNA kit (Gentra, Minneapolis, MN) or the AllPrep DNA/RNA Mini Kit (Qiagen Sciences, Maryland, USA) with slight modifications (as in Ward 2008).
Label Cy5
Label protocol Hybridization targets were prepared by digesting environmental DNA with a four cutter restriction enzyme. The digested environmental DNA were labeled with amino-allyl-dUTP (Ambion) during linear amplification using random octamers and a Klenow polymerase (Applied Biosystems). The reaction contained 3.96 mM d(AGC)TP, 0.44 mM dTTP, and 4.84 mM dUaa and was amplified for 3 h at 37°C. The Klenow product was purified and conjugated with Cy3. The Cy3-labelled target (1000 ng) was combined with 2x hybridization buffer (1x final concentration; Agilent) and 0.25 pmol of a Cy5-labelled complementary 20-mer standard oligonucleotide and incubated at 95°C for 5 min before being cooled to room temperature.
 
Channel 2
Source name Total DNA from water column, Arabian Sea station 1, 175m
Organism marine metagenome
Characteristics location: Arabian Sea Station 1
depth: 175 m
sample type: environmental
Extracted molecule genomic DNA
Extraction protocol DNA was extracted from the filters using the PureGene DNA kit (Gentra, Minneapolis, MN) or the AllPrep DNA/RNA Mini Kit (Qiagen Sciences, Maryland, USA) with slight modifications (as in Ward 2008).
Label Cy3
Label protocol Hybridization targets were prepared by digesting environmental DNA with a four cutter restriction enzyme. The digested environmental DNA were labeled with amino-allyl-dUTP (Ambion) during linear amplification using random octamers and a Klenow polymerase (Applied Biosystems). The reaction contained 3.96 mM d(AGC)TP, 0.44 mM dTTP, and 4.84 mM dUaa and was amplified for 3 h at 37°C. The Klenow product was purified and conjugated with Cy3. The Cy3-labelled target (1000 ng) was combined with 2x hybridization buffer (1x final concentration; Agilent) and 0.25 pmol of a Cy5-labelled complementary 20-mer standard oligonucleotide and incubated at 95°C for 5 min before being cooled to room temperature.
 
 
Hybridization protocol Samples were hybridized to duplicate arrays by overnight incubation at 64°C and washed.
Scan protocol The arrays were scanned with a laser scanner (Molecular Devices 4200).
Images were analyzed with GenePix Pro 6.0 software (Molecular Devices).
Description AS1.175m
This Sample represents 2 replicates.
DNA extracted from sterivex filter used to create 2 targets to run on duplicate arrays.
Data processing Quality control was employed to remove noise by eliminating features 1) in which the Cy3 signal was not at least twice the value of the control probes, and 2) in which the Cy3/Cy5 could not pass a Z test to find out the outlier values. The Z test was performed by calculating Zi = ri/(s/square root of 3), i = 1, 2, or 3, where ri represents Cy3/Cy5, s the standard deviation of the triplicate Cy3/Cy5 values. A feature is considered as an outlier if the Z is greater than 1.9 (CI = 80%).
Cy3/Cy5 fluorescence intensities were standardized to the highest Cy3/Cy5 fluorescence across the AOA probe set (normalized fluorescence ratio; FRN).
 
Submission date May 12, 2013
Last update date Jun 20, 2013
Contact name Xuefeng Peng
Organization name Princeton University
Department Geosciences
Street address Guyot Hall
City Princeton
State/province NJ
ZIP/Postal code 08544
Country USA
 
Platform ID GPL17151
Series (1)
GSE46851 Community composition of ammonia-oxidizing archaea from surface and anoxic depths of oceanic oxygen minimum zones

Data table header descriptions
ID_REF
VALUE Log2 normalized Cy3/C5 ratio
PRE_VALUE Normalized Cy3/C5 ratio

Data table
ID_REF VALUE PRE_VALUE
AOA1 -4.9658 0.032
AOA2 -4.5735 0.042
AOA3 -2.4580 0.182
AOA4 -1.0291 0.490
AOA5 -5.7959 0.018
AOA6 -5.5064 0.022
AOA7 -4.0831 0.059
AOA8 -4.7179 0.038
AOA9 -2.4112 0.188
AOA10 -2.2109 0.216
AOA11 -3.4901 0.089
AOA12 -2.4112 0.188
AOA13 -4.5395 0.043
AOA14 -3.2379 0.106
AOA15 -1.8890 0.270
AOA16 -2.9771 0.127
AOA17 0.0000 1.000
AOA18 -4.9658 0.032
AOA19 -1.6349 0.322
AOA20 -4.2379 0.053

Total number of rows: 31

Table truncated, full table size <1 Kbytes.




Supplementary file Size Download File type/resource
GSM1139165_BC014-19-3_Irregular.gpr.gz 148.4 Kb (ftp)(http) GPR
GSM1139165_BC014-19-4_Irregular.gpr.gz 153.6 Kb (ftp)(http) GPR
GSM1139165_BC014_19_3NewFilterS-Irbbw.xlsx.gz 715.5 Kb (ftp)(http) XLSX
GSM1139165_BC014_19_4NewFilterS-Irbbw.xlsx.gz 726.5 Kb (ftp)(http) XLSX
Processed data included within Sample table
Processed data provided as supplementary file

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