Almost all animals are young adults between 60 and 100 days of age (Table 1, minimum age is 60 and maximum age is 118 days). We measured expression in conventional inbred strains, BXD recombinant inbred (RI) strains, and reciprocal F1s between C57BL/6J and DBA/2J.
- -BXD strains:
- -Sample Processing: Dr. XiangDi Wang (UTHSC) and Becky King (Emory) were involved in the retinal extractions and isolation of RNA. The Affymetrix arrays were run by two different research cores: the Molecular Resource Center at UTHSC (Dr. William Taylor Director) and the Integrated Genomics Core at Emory University by Robert B Isett (Dr. Michael E. Zwick, Director). In a separate set of experiments we could not detect any significant difference in the arrays run at UTHSC or at Emory University.
- -Replication, sex, and sample balance: Our goal was to obtain data for independent biological sample pools from both sexes for most lines of mice. The four batches of arrays included in this final data set, collectively represent a reasonably well-balanced sample of males and females, in general without within-strain-by-sex replication.
diff --git a/general/datasets/DoDTATRCRetExMoGene2_0315/notes.rtf b/general/datasets/DoDTATRCRetExMoGene2_0315/notes.rtf deleted file mode 100644 index cb14c74..0000000 --- a/general/datasets/DoDTATRCRetExMoGene2_0315/notes.rtf +++ /dev/null @@ -1,10 +0,0 @@ -Other Data Sets Users of these mouse retina data may also find the following complementary resources useful:
- -Affymetrix Mouse Gene 2.0 ST Array: These expression arrays have been designed with a median of 22 unique probes per transcript. Each unique probe is 25 bases in length, which means that the array measures a median of 550 bases per transcript. The arrays provide comprehensive transcriptome coverage with over 30,000 coding and non-coding transcripts. In addition there is coverage for 592 microRNAs. For some arrays the RNA was pooled from two retinas and for other arrays were run on a single retina.
diff --git a/general/datasets/DoDTATRCRetExMoGene2_0315/summary.rtf b/general/datasets/DoDTATRCRetExMoGene2_0315/summary.rtf deleted file mode 100644 index b6cf222..0000000 --- a/general/datasets/DoDTATRCRetExMoGene2_0315/summary.rtf +++ /dev/null @@ -1,5 +0,0 @@ -DoD TATRC Retina Dataset Affymetrix MouseGene 2.0 ST Array (____ 2015) RMA analysis and scaling by Arthur Centeno. This data set consists of 75 BXD strains, C57BL/6J, DBA/2J, both reciprocal F1s, and BALB/cByJ. A total of 80 strains were quantified. The data are now open and available for analysis.
- -This is RMA expression data that has been normalized using what we call a 2z+8 scale, but without special correction for batch effects. The data for each strain was computed as the mean of four samples per strain. Expression values on a log2 scale range from 6.25 to 18.08 (11.83 units), a nominal range of approximately 3600-fold. After taking the log2 of the original non-logged expression estimates, we convert data within an array to a z score. We then multiply the z score by 2. Finally, we add 8 units to ensure that no values are negative. The result is a scale with a mean expression of the probes on the array of 8 units and a standard deviation of 2 units. A two-fold difference in expression is equivalent roughly to 1 unit on this scale.
- -The lowest level of expression is 6.254 for ILMN_2747167 (Arhgap11a) from HEI Retina Illumina V6.2 (April 2010) RankInv **. Lowest single data about 5.842. The highest level of expression is 18.077 for ILMN_2516699 (Ubb). Highest single value is about 18.934.
diff --git a/general/datasets/DoDTATRCRetExMoGene2_0315/tissue.rtf b/general/datasets/DoDTATRCRetExMoGene2_0315/tissue.rtf deleted file mode 100644 index 87eb144..0000000 --- a/general/datasets/DoDTATRCRetExMoGene2_0315/tissue.rtf +++ /dev/null @@ -1,13 +0,0 @@ -Tissue preparation protocol. Animal were killed by rapid cervical dislocation. Retinas were removed immediately and placed in 1 ml of 160 U/ml Ribolock for 1 min at room temperature. Dissecting and preparing eyes for RNA extraction Retinas for RNA removed from the eye and placed in Hank’s Balanced Salt solution with RiboLock (Thermo Scientific RiboLock RNase #EO0381 40U/µl 2500U) and processed per manufacturer’s instructions (in brief form below).
- -1) Sample collection for RNA isolation.
- -2) Quickly remove the retinas with clean curved forceps after cervical dislocation of the mouse.
- -3) Put each retina in 1 ml of 160 U/ml Ribolock for 1 min in RT.
- -4) Move the retina to another tube with 50µl Ribolock, store in -80°C.
- -5) The RNA was isolated using a QiaCube and the in column DNAse procedure.
- -Quality Control: All RNA samples were checked for quality before running microarrays. The samples were analyzed using the Agilent 2100 Bioanalyzer. The RNA integrity values for each sample are presented in Table 1 below.
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