Description
This track shows the transcript map of signal intensity (estimating RNA
abundance) for the following, hybridized to the Affymetrix ENCODE
oligonucleotide microarray:
- human neutrophil (PMN) total RNA (10 biological samples from different individuals)
- human placental Poly(A)+ RNA (3 biological replicates)
- total RNA from human NB4 cells (4 biological replicates), each sample
divided into three parts and treated as follows: untreated, treated with
retinoic acid (RA), and treated with 12-O-tetradecanoylphorbol-13 acetate (TPA)
(three out of the four original samples). Total RNA was extracted from each
treated sample and applied to arrays in duplicate (2 technical replicates).
The human NB4 cell can be made to differentiate towards either monocytes (by
treatment with TPA) or neutrophils (by treatment with RA). See Kluger
et al., 2004 in the References section for more details about the
differentiation of hematopoietic cells.
This array has 25-mer oligonucleotide probes tiled
approximately every 22 bp, covering all the non-repetitive DNA sequence
of the ENCODE regions. The transcript map is a combined signal for both
strands of DNA. This is derived from the number of different biological
samples indicated above, each with at least two technical replicates.
See the following NCBI Gene Expression Omnibus (GEO) accessions for details
of experimental protocols:
- ENCODE Transcript Mapping for Human Neutrophil (PMN) Total RNA:
GSE2678
- ENCODE Transcript Mapping for Human Placental Poly(A)+ RNA:
GSE2671
- ENCODE Transcript Mapping for Total RNA from Human NB4 Cells untreated,
treated with RA, and treated with TPA:
GSE2679
Display Conventions and Configuration
This annotation follows the display conventions for composite
"wiggle" tracks. The subtracks within this annotation
may be configured in a variety of ways to highlight different aspects of the
displayed data. The graphical configuration options are shown at the top of
the track description page, followed by a list of subtracks. To display only
selected subtracks, uncheck the boxes next to the tracks you wish to hide.
For more information about the graphical configuration options, click the
Graph
configuration help link.
Color differences among the subtracks are arbitrary. They provide a
visual cue for distinguishing between the different data samples.
Methods
The data from biological & technical replicates were
quantile-normalized to each other and then median scaled
to 25. Using a 101 bp sliding window centered on each
oligonucleotide probe, a signal map estimating RNA
abundance was generated by computing the pseudomedian
signal of all PM-MM pairs (median of pairwise PM-MM
averages) within the window, including replicates.
Verification
Independent biological replicates (as indicated above) were generated,
and each was hybridized to at least two different arrays
(technical replicates). Transcribed regions were then identified using a
signal threshold of 90 percentile of signal intensities, as well as a maximum
gap of 50 bp and a minimum run of 50 bp (between oligonucleotide positions).
Transcribed regions, as determined by individual biological samples, were
compared to ensure significant overlap.
Credits
These data were generated and analyzed by the Yale/Affymetrix
collaboration between the labs of Michael Snyder, Mark Gerstein and
Sherman Weissman at Yale University and Tom Gingeras at Affymetrix.
References
Bertone P, Stolc V, Royce TE, Rozowsky JS, Urban AE, Zhu X,
Rinn JL, Tongprasit W, Samanta M, Weissman S et al.
Global identification of human transcribed sequences with
genome tiling arrays.
Science. 2004 Dec 24;306(5705):2242-6.
Cheng J, Kapranov P, Drenkow J, Dike S, Brubaker S, Patel S,
Long J, Stern D, Tammana H, Helt G et al.
Transcriptional maps of 10 human chromosomes at 5-nucleotide
resolution.
Science. 2005 May 20;308(5725):1149-54.
Kapranov P, Cawley SE, Drenkow J, Bekiranov S, Strausberg RL, Fodor SP,
Gingeras TR.
Large-scale transcriptional activity in chromosomes 21 and
22.
Science. 2002 May 3;296(5569):916-9.
Kluger Y, Tuck DP, Chang JT, Nakayama Y, Poddar R, Kohya N,
Lian Z, Ben Nasr A, Halaban HR, Krause DS et al.
Lineage specificity of gene expression patterns.
Proc Natl Acad Sci U S A. 2004 April 27;101(17):6508-13.
Rinn JL, Euskirchen G, Bertone P, Martone R, Luscombe NM,
Hartman S, Harrison PM, Nelson FK, Miller P, Gerstein M et al.
The transcriptional activity of human Chromosome 22.
Genes Dev. 2003 Feb 15;17(4):529-40.
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