From ea46f42ee640928b92947bfb204c41a482d80937 Mon Sep 17 00:00:00 2001 From: root Date: Tue, 8 May 2012 18:39:56 -0500 Subject: Add all the source codes into the github. --- web/dbdoc/Eye_M2_0608_R.html | 486 +++++++++++++++++++++++++++++++++++++++++++ 1 file changed, 486 insertions(+) create mode 100755 web/dbdoc/Eye_M2_0608_R.html (limited to 'web/dbdoc/Eye_M2_0608_R.html') diff --git a/web/dbdoc/Eye_M2_0608_R.html b/web/dbdoc/Eye_M2_0608_R.html new file mode 100755 index 00000000..cc05b67e --- /dev/null +++ b/web/dbdoc/Eye_M2_0608_R.html @@ -0,0 +1,486 @@ + +HEIMED M430 Microarray Eye RMA September06 / GN + + + + + + + + + + + + + + + + + +
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+ +Hamilton Eye Institute Mouse Eye M430v2 (Sept06) RMA Data Set modify this page

Accession number: GN162

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    Summary:

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+RECOMMENDED EYE DATA SET. The HEIMED September 2006 RMA fixed data set provides estimates of mRNA expression in whole eyes of 84 lines of young adult mice generated using approximately 175 Affymetrix M430 2.0 arrays. This data set corrects probable errors in strain assignment that affected strains BXD2, BXD31, BXD89, and A/J. Data were generated at UTHSC with support from a grant from Dr. Barrett Haik, Director of the Hamilton Eye Institute (HEI). We used pooled RNA samples, usually two independent pools; one male, one female, for each strain. This data set was processed using the RMA protocol. To simplify comparison among different transforms, RMA values of each array have been adjusted to an average expression of 8 units and a standard deviation of 2 units. Values range from a low of 4.78 (Tcf15, probe set 1420281_at) to a high of 15.47 (crystallin gamma C, Crygc, probe set 1422674_s_at). Even probe sets with comparatively low expression can provide reliable data. For example probe set 1440397_at (Cacna2d1) has expression of only 5.5, but is associated with a cis QTL with an LRS of 79. This indicates a high signal to noise ratio and the detection of significant strain variation of the correct transcript.

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An additional set of strains will be added in early fall 2008 to complete the HEIMED data set. We will be adding data for the following strains: + +

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  1. Three additional BXD strains (67 total). The three new strains are BXD56, BXD71, and BXD99. +
  2. More arrays for the parental strains, C57BL/6J and DBA/2J, and their reciprocal F1s (n = 4) +
  3. Ten new common strains of mice. The Mouse diversity panel will include a total of 29 strains (n = 27 strains plus B6 and D2 +
  4. Seven KO lines (Rpe65, Nyx (NOB), Gabbr1, Gnb1, Gabra1, Gpr19, and Clcn3) +
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Users of these mouse eye data may also find the NEIBank collection of ESTs and SAGE data of substantial utility. +

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    About the cases used to generate this set of data:

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+We used a set of 64 BXD recombinant inbred strains, 18 conventional inbred strains including C57BL/6J (B6) and DBA/2J (D2), and their reciprocal F1s. BXD strains were generated by crossing C57BL/6J with DBA/2J. The BXDs are particularly useful for systems genetics because both parental strains have been sequenced (8x coverage of B6 and 1.5x coverage of D2). Physical maps in WebQTL incorporate approximately 2 million B6 vs D2 SNPs from Celera Genomics and from the Perlegen-NIEHS sequencing effort. BXD1 through BXD32 were bred by Benjamin A. Taylor starting in the late 1970s. BXD33 through 42 were bred by Taylor in the 1990s. These strains are available from The Jackson Laboratory. + +

Please note that BXD24/TyJ (JAX stock number 000031) used in this study is also known as BXD24b/TyJ and has nearly complete retinal degeneration. BXD24a/TyJ, a 1988 F80 stock that has now been rederived, does not have retinal degeneration (stock number 005243) and is an ideal coisogenic control. + +

BXD43 through BXD99 were bred by Lu Lu, Jeremy Peirce, Lee M. Silver, and Robert W. Williams in the late 1990s and early 2000s using advanced intercross progeny (Peirce et al. 2004). Many of the 50 new BXD strains are available from Lu Lu and colleagues.

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Mouse Diversity Panel (MDP). In addition to the BXD strains, we have profiled a MDP consisting 18 inbred strains and a pair of reciprocal F1 hybrids; B6D2F1 and D2B6F1. These strains were selected for several reasons: + +

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  • genetic and phenotypic diversity, including use by the Phenome Project +
  • their use in making genetic reference populations including recombinant inbred strains, cosomic strains, congenic and recombinant congenic strains +
  • their use by the Complex Trait Consortium to make the Collaborative Cross (Nairobi/Wellcome, Oak Ridge/DOE, and Perth/UWA) +
  • genome sequence data from three sources (NHGRI, Celera, and Perlegen-NIEHS) +
  • availability from The Jackson Laboratory +
+ +

All eight parents of the Collaborative Cross (129S1/SvImJ, A/J, C57BL/6J, CAST/EiJ, NOD/LtJ, NZO/HlLtJ, PWK/PhJ, and WSB/EiJ) have been included in the MDP. Twelve MDP strains have been sequenced or resequenced by Perlegen for the NIEHS (see the GeneNetwork SNP Browser for data, details, and a link to Perlegen Inc excellent data resources and browser). + +

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  1. 129S1/SvImJ +
        Collaborative Cross strain sequenced by NIEHS; background for many knockouts (R1 ES cell line); Phenome Project A list. This strain (JAX No 002448, aka 129S1/Sv-++Kitl/+) also carries hypopigmentation mutations (white bellied chinchilla) of the tyrosinase gene on Chr 7 and a mutant allele of the steel (Kitl) gene. (JAX Stock Number: 002448) + +
  2. A/J +
        Collaborative Cross strain sequenced by Perlegen/NIEHS; parent of the AXB/BXA panel. A tyrosinase-negative albino (c) mutant + + +
  3. BALB/cByJ +
         Sequenced by NIEHS; maternal parent of the CXB panel; Phenome Project A list. A tyrosinase-negative albino (c) mutant. (JAX Stock Number: 001026) + +
  4. BXSB/MpJ +
         A strain with interesting autoimmune disease associated with glomerulonephritis. (JAX Stock Number: 000740) + +
  5. C3H/HeJ +
        Sequenced by Perlegen/NIEHS; paternal parent of the BXH panel; Phenome Project A list. Important to note for this Eye dataset, C3H/HeJ is a Pdeb6 mutant with near total photoreceptor loss at as early as postnatal day 30. + +
  6. C57BL/6J +
        Sequenced by NIH/NHGRI; parental strain of AXB/BXA, BXD, and BXH; Phenome Project A list + +
  7. CAST/EiJ (please note in an early data release, we listed CAST/Ei and CAST/EiJ as two different strains) +
        Collaborative Cross strain sequenced by NIEHS; Phenome Project A list (JAX Stock Number: 000928). + +
  8. DBA/2J +
        Sequenced by Perlegen/NIEHS and Celera; paternal parent of the BXD panel; Phenome Project A list + +
  9. FVB/NJ +
        Sequenced by Perlegen/NIEHS and Celera; most common strain used to make transgenic mice due to large and easily injected oocyte; Phenome Project A list (JAX Stock Number: 001800). + +
  10. KK/HlJ +
        Sequenced by Perlegen/NIEHS + +
  11. LG/J +
        Paternal parent of the LGXSM panel + +
  12. MOLF/EiJ +
        A wild strain derived from the M. musculus molossinus subspecies of mice that has retinal degeneration affecting photoreceptors. There appears to have been some genetic contamination or admixture of this strain with conventional inbred strains in the very recent past (F. Pardo, personal communication to RWW, August 2006) (JAX Stock Number: 000550). + +
  13. NOD/LtJ +
        Collaborative Cross strain sequenced by NIEHS; Phenome Project B list; diabetic + +
  14. NZO/HlLtJ +
        Collaborative Cross strain + +
  15. PWD/PhJ +
        Sequenced by Perlegen/NIEHS; parental strain for a consomic set by Forjet and colleagues + +
  16. PWK/PhJ +
        Collaborative Cross strain; Phenome Project D list + +
  17. WSB/EiJ
        Collaborative Cross strain sequenced by NIEHS; Phenome Project C list (JAX Stock Number: 001145). + +
  18. B6D2F1 and D2B6F1, aka F1 in some graphs and tables +
    F1 hybrids generated by crossing C57BL/6J with DBA/2J. These reciprocal F1 can be used to detect some imprinted genes. +
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    About the tissue used to generate this set of data:

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Tissue preparation protocol. Animal were killed by rapid cervical dislocation. Eyes were removed immediately and placed in RNAlater at room temperature. Usually six eyes from animals with a common sex, age, and strain were stored in a single tube. + +

Each array was hybridized with a pool of RNA from 4 to 8 eyes from 2 to 4 animals. RNA was extracted at UTHSC by Zhiping Jia. If tissue was saved for RNA extraction at a later time, eyes were placed directly in RNAlater (Ambion, Inc.) and treated per the manufacturer’s directions. If eyes were used for immediate RNA extraction then we proceeded immediately to the next steps.

+ +Dissecting and preparing eyes for RNA extraction +

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  1. Place eyes for RNA extraction in RNA STAT-60 (Tel-Test Inc.) and process per manufacturer’s instructions (in brief form below). +
  2. Store RNA in 75% ethanol at –80 deg. C until use. +
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Total RNA was extracted with RNA STAT-60 (Tel-Test Inc.) according to the manufacturer's instructions. Briefly we: +

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  1. homogenize tissue samples in the RNA STAT-60 (1 ml/50 to 100 mg tissue) +
  2. allowed the homogenate to stand for 5 min at room temperature +
  3. added 0.2 ml of chloroform per 1 ml RNA STAT-60 +
  4. shook the sample vigorously for 15 sec and let the sample sit at room temperature for 3 min +
  5. centrifuged at 12,000 G for 15 min +
  6. transfered the aqueous phase to a fresh tube +
  7. added 0.5 ml of isopropanol per 1 ml RNA STAT-60 +
  8. vortexed and allowed sample to stand at room temperature for 5-10 min +
  9. centrifuged at 12,000 G for 10-15 min +
  10. removed the supernatant and washed the RNA pellet with 75% ethanol +
  11. stored the pellet in 75% ethanol at -80 deg C until use +
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Sample Processing. Samples were processed in the VA Medical Center, Memphis, Rheumatology Disease Research Core Center lead by Drs. John Stuart and Weikuan Gu. All processing steps were performed by Dr. Yan Jiao. In brief, samples were purified using a standard sodium acetate in alcohol method (recommended by Affymetrix). The RNA quality was checked using a 1% agarose gel. The 18S and 28S bands had to be clear and the 28S band had to be more prominent. RNA concentation was measured using a spectrophotometer. The 260/280 ratios had to be greater than 1.7, and the majority were 1.8 or higher. We used a total of 8 micrograms of RNA as starting amount for cDNA synthesis using a standard Eberwine T7 polymerase method (Superscript II RT, Invitrogen Inc., Affy Part No 900431, GeneChip Expression 3' Amplification One-Cyle cDNA Synthesis Kit). The Affymetrix IVT labeling kit (Affy 900449) was used to generate labeled cRNA. At this point the cRNA was evaluated again using both the 260/280 ratio (values of 2.0 or above were acceptable) and 1% agarose gel inspection of the product (a size range from 200 to 7000 bp is considered suitable for use). We used 45 micrograms of labeled cRNA for fragmentation. Those samples that passed both QC steps (<10% usually fail) were then sheared using a fragmentation buffer included in the Affymetrix GeneChip Sample Cleanup Module (Part No.900371). After fragmentation, samples were either stored at -80 deg. centrigrade until use (one third roughly) or were immediately used for hybridization. + +

Dealing with Ocular Pigmentation: Variable ocular pigmentation is a potential confound in a study of the whole eye transcriptome. Even the most careful RNA preparations taken from brown and beige colored mice tend to have faint residual pigmentation that affects hybridization signal. The key determinant of this interesting effect is the Tyrp1 (brown) locus on Chr 4 at about 80 Mb. Loci on Chr 4 that map at this location should be considered with skepticism and reviewed carefully. To address this problem Yan Jiao purified total RNA a second time using the Qiagen RNeasy MinElute Cleanup Kit (Cat No. 74204). This was done for 8 colored samples (R2534E2, R2578E2, R1441E2, R2537E2, R2536E2, R2589E2, R2539E2) in the first batch of arrays (the November 05 data set) of which 5 were finally included in this data set (cases in which the strain ID is labeled with asterisks in the table). This same protocol was used for all samples in the second and third batches added in April 2006 and September 2006. + +

Replication and Sample Balance: Our goal is to obtain data for independent biological sample pools from at least one of sample from each sex for all BXD strains. The three batches of arrays included in this September data set, collectively represent a reasonably well balanced sample of males and females belonging to 84 strains, but without within-strain-by-sex replication. Three strains are represented only by male sample pools (A/J, BXD29, BXD48). One strain is represented only by a female pool sample (BXD89). Please use the probe sets 1427262_at (Xist, high in females) and probe set 1426438_at (Ddx3y, high in males) as quantitative surrogates for the sex balance in this data set. + +

Batch Structure: This data set consists of a three batches: the original batch that makes up the November 2005 data set (n = XXX), a second batch of 63 arrays (R0857E through R2649E, and R2682E through R2742E, non-consecutive identifiers) run in January 2006 by Dr. Yan Jiao; and a third batch of 41 arrays (XXXX through YYYYY) run in August 2006 by Dr. Yan Jiao. The arrays in the first batches are from different lots. +All arrays in the first batch were from Lot YYYYY (expiration date XX.YY.ZZ). +All arrays in the second batch were from Lot 4016879 (expiration date 12.28.06). +All NN arrays in the third batch were from Lot XXXXX (expiration date XX.YY.ZZ). + +We started working with a total of approximately 190 (???) arrays that passed initial crude quality control based on RNA quality and initial Affymetrix report file information such as 3'/5' ratio, scale factor, and percent present calls. A total of 177 (???) arrays were finally approved for inclusion in this September 2006 data set. The complex normalization procedure is described below. + +

The table below summarizes information on strain, age, sex, original CEL filename, Affymetrix quality control values, and source of mice. Columns labeled "PDNN 2Z outlier" and "RMA 2Z outlier" list the fraction of probe sets with values that deviated more than 2 z units from the mean. Scale factor, background average, present, absent, marginal and 3'/5' ratios for actin and Gapdh were collated from the Affymetrix Report (RPT) files. + +

IN PROGRESS: PLEASE NOTE THAT THIS TABLE IS NOW BEING UPDATED TO INCLUDE BATCHES 2 and 3 OF EARLY and MID 2006. + +

+ + + ++++ + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + + +
ID +

tube ID

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group_type

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 Strain

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age

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 Sex

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original

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CEL

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filename

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PDNN

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2Z

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outlier

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RMA

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2Z

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outlier

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scale

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factor

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background

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average

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present

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absent

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marginal

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AFFX-b-

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ActinMur(3'/5')

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AFFX-

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GapdhMur(3'/5')

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Source

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1R2533E1GDP129S1/SvImJ60MR2533E.CEL0.0250.0282.119457.90%40.50%1.60%1.370.78UTM RW
2R2595E1GDP129S1/SvImJ59FR2595E.CEL0.0330.0361.7911561.00%37.50%1.50%1.460.77UTM RW
3R0754E2GDPA/J60MR0754E.CEL0.0270.032.728659.80%38.70%1.50%1.360.76JAX
4R2546E1GDPA/J66FR2545E.CEL0.0240.0291.999658.60%39.70%1.70%1.470.78UTM RW
5R2601E1GDP BXDB6D2F173FR2601E.CEL0.0070.0082.559258.90%39.60%1.50%1.440.78UTM RW
6R2602E1GDP BXDB6D2F173MR2602E.CEL0.0030.0082.68459.70%38.80%1.50%1.370.78UTM RW
7R1672E1GDPBALB/cByJ83MR1672E.CEL0.0430.0392.2211159.90%38.60%1.50%1.260.8JAX
8R1676E1GDPBALB/cByJ83FR1676E.CEL0.0830.0852.699858.90%39.60%1.50%1.460.74JAX
9R2704EBXDBXD159FR2704E.CEL0.0290.032.066139.6156.60%41.90%1.50%1.310.81GU
10R2581E1BXDBXD1165FR2581E.CEL0.0090.0211.948962.10%36.40%1.60%1.550.81UTM RW
11R2612EBXDBXD1170MR2612E.CEL0.1010.1121.83142.0358.20%40.50%1.40%1.780.81GU
12R2543E1BXDBXD1263MR2543E.CEL0.0180.0171.6111858.60%39.90%1.60%1.430.77UTM RW
13R2742EBXDBXD1271FR2742E.CEL0.0730.0772.127134.1457.00%41.60%1.40%1.640.78GU
14R2586E1BXDBXD1360FR2586E.CEL0.2590.2582.017456.40%42.00%1.60%2.853.81Glenn
15R877EBXDBXD1376MR877E.CEL0.0260.0671.558125.6361.20%37.50%1.20%1.420.81GU
16R2557E1BXDBXD1460FR2557E.CEL0.0120.0271.839962.50%36.10%1.40%1.310.78Glenn
17R1128EBXDBXD1465MR1128E.CEL0.0370.0382.366118.3957.30%41.30%1.40%1.450.81GU
18R2716EBXDBXD1560MR2716E.CEL0.0350.0372.015150.8356.40%42.10%1.60%1.420.81GU
19R2567E1BXDBXD1660MR2567E.CEL0.0480.0582.248256.70%41.60%1.70%1.370.75Glenn
20R2711EBXDBXD1661FR2711E.CEL0.0320.0211.953118.5359.00%39.60%1.50%1.450.8GU
21R2559E1BXDBXD1859MR2559E.CEL0.010.0121.6510460.80%37.70%1.50%1.270.78Glenn
22R2720EBXDBXD1859FR2720E.CEL0.0140.0192.3299.9359.50%39.00%1.50%1.330.77GU
23R2560E1BXDBXD1960FR2560E.CEL0.0090.0121.799860.90%37.50%1.60%1.350.8Glenn
24R2713EBXDBXD1960MR2713E.CEL0.0550.0211.67120.8260.20%38.30%1.50%1.450.8GU
25R2597E1BXDBXD261MR2597E.CEL0.0050.0122.379460.30%38.30%1.50%1.340.77Glenn
26R1231EBXDBXD264FR1231E.CEL0.0440.0372.197138.7357.30%41.30%1.40%1.410.77GU
27R2584E1BXDBXD2059FR2584E.CEL0.0110.0172.078459.30%39.10%1.60%1.40.76Glenn
28R2731EBXDBXD2060MR2731E.CEL0.0170.0191.82514759.00%39.50%1.50%1.40.8GU
29R2541E2BXDBXD2161MR2541E2.CEL0.0490.0842.6312556.00%42.40%1.50%1.290.78UTM RW
30R2702EBXDBXD2159FR2702E.CEL0.0090.0081.811128.6559.40%39.10%1.40%1.260.8GU
31R2553E1BXDBXD2258FR2553E.CEL0.0040.011.9511159.90%38.50%1.50%1.280.76Glenn
32R2700EBXDBXD2259MR2700E.CEL0.010.0151.858102.9661.50%37.10%1.30%1.480.79GU
33R2558E1BXDBXD2360FR2558E-2.CEL0.0180.0271.9111559.90%38.80%1.40%1.20.82Glenn
34R1086EBXDBXD2355MR1086E.CEL0.0430.0342.233125.0558.60%39.90%1.50%1.430.77GU
35R2589E2BXDBXD2459MR2589E2.CEL0.1320.1762.6111257.50%40.90%1.60%1.240.8Glenn
36R2719EBXDBXD24123FR2719E.CEL0.1120.1111.47140.3861.50%37.20%1.30%1.380.79GU
37R2573E1BXDBXD2567FR2573E-2.CEL0.0550.0633.157257.90%40.70%1.40%1.770.97UAB
38R2683EBXDBXD2558MR2683E.CEL0.0680.0681.777115.6458.30%40.30%1.40%2.010.79GU
39R2703EBXDBXD2760FR2703E.CEL0.0080.0121.263134.7862.60%36.10%1.40%1.440.78GU
40R2721EBXDBXD2860MR2721E.CEL0.040.0482.065157.3956.10%42.40%1.50%1.310.81GU
41R2562E1BXDBXD2960MR2562E.CEL0.0070.011.6511659.90%38.40%1.70%1.370.79Glenn
42R2598E1BXDBXD3161MR2598E.CEL0.0060.0131.9910660.90%37.60%1.50%1.270.78UTM RW
43R1258EBXDBXD3157FR1258E.CEL0.0370.0362.063117.0959.00%39.50%1.50%1.540.78GU
44R2563E1BXDBXD3263FR2563E.CEL0.0230.0251.5510261.90%36.70%1.40%1.50.8UTM RW
45R1216EBXDBXD3276MR1216E.CEL0.050.0492.23111.9958.80%39.80%1.40%1.350.79GU
46R2542E1BXDBXD3367FR2542E.CEL0.0580.0622.139756.50%41.80%1.60%1.910.93UTM RW
47R857EBXDBXD3377MR857E.CEL0.0780.1081.737113.9861.90%36.70%1.30%1.60.77GU
48R2585E1BXDBXD3460MR2585E.CEL0.0240.0322.647558.30%40.00%1.70%1.250.77Glenn
49R1451EBXDBXD3461FR1451E.CEL0.010.0091.843140.0559.00%39.50%1.50%1.420.81GU
50R2532E1BXDBXD3862MR2532E.CEL0.0020.0062.049459.80%38.70%1.50%1.370.8UTM RW
51R2710EBXDBXD3855FR2710E.CEL0.0330.0312.112122.158.80%39.80%1.40%1.370.78GU
52R2574E1BXDBXD3970FR2574E.CEL0.0030.0081.989161.20%37.30%1.50%1.390.78UTM RW
53R2695EBXDBXD3959MR2695E.CEL0.0180.0161.638122.760.80%37.80%1.50%1.420.8GU
54R2590E1BXDBXD4060MR2590E.CEL0.0070.0122.717759.10%39.30%1.50%1.40.77Glenn
55R2699EBXDBXD4059FR2699E.CEL0.0140.0151.827105.2361.70%36.90%1.40%1.420.81GU
56R2596E1BXDBXD4259MR2596E.CEL0.0160.032.6310859.00%39.60%1.50%1.240.8Glenn
57R2696EBXDBXD4258FR2696E.CEL0.010.0171.622118.9562.00%36.60%1.50%1.530.79GU
58R2605E1BXDBXD4379MR2607E.CEL0.0060.011.8213160.50%38.20%1.30%1.320.8UTM RW
59R994EBXDBXD4360FR994E.CEL0.0130.0141.966113.1260.80%37.80%1.40%1.660.8GU
60R2594E1BXDBXD4463FR2594E.CEL0.0140.0241.7711759.80%38.80%1.40%1.350.85UTM RW
61R2610EBXDBXD4468MR2610E.CEL0.0130.0091.814142.9159.00%39.50%1.50%1.350.8GU
62R2592E1BXDBXD4562MR2592E.CEL0.0050.0111.8510660.10%38.60%1.30%1.430.85UTM RW
63R2732EBXDBXD4563FR2732E.CEL0.0390.0362.154122.4556.50%42.10%1.40%1.80.83GU
64R2606E1BXDBXD4878MR2606E.CEL0.0070.0152.5610658.90%39.70%1.40%1.350.83UTM RW
65R967EBXDBXD4864FR967E.CEL0.1010.0521.948130.9557.30%41.20%1.50%1.630.81GU
66R2591E1BXDBXD560FR2591E.CEL0.0520.0141.713658.50%40.00%1.50%1.330.78Glenn
67R2714EBXDBXD558MR2714E.CEL0.0470.0141.404144.3560.60%37.90%1.50%1.430.79GU
68R2603E1BXDBXD5166FR2603E.CEL0.0070.022.4911557.70%40.80%1.50%1.240.79UTM RW
69R1042EBXDBXD5162MR1042E.CEL0.0280.0272.352104.1258.70%39.90%1.40%1.530.82GU
70R2690EBXDBXD5565MR2690E.CEL0.0810.0671.887164.0156.10%42.30%1.60%1.430.8GU
71R2570E1BXDBXD665FR2570E.CEL0.0130.0171.998758.50%40.00%1.50%1.460.76UTM RW
72R2694EBXDBXD658MR2694E.CEL0.0120.0181.98397.2361.60%37.10%1.30%1.390.82GU
73R2534E2BXDBXD6170FR2534E2.CEL0.030.0582.4711857.90%40.60%1.50%1.420.79UTM RW
74R2684EBXDBXD6162MR2684E.CEL0.0310.0322.01131.0357.00%41.50%1.50%1.340.78GU
75R2611E1BXDBXD6468MR2611E.CEL0.0670.0682.299258.00%40.50%1.50%1.571.06UTM RW
76R943E-2BXDBXD6456FR943E-2.CEL0.0240.0211.591141.3460.10%38.40%1.50%1.320.76GU
77R2583E1BXDBXD6560MR2583E.CEL0.0270.032.497056.90%41.50%1.60%1.671.01UTM RW
78R2689EBXDBXD6563FR2689E.CEL0.0080.0081.721142.4459.90%38.60%1.50%1.380.76GU
79R2536E2BXDBXD6664FR2536E2.CEL0.0670.1392.7410956.10%42.30%1.70%1.280.79UTM RW
80R1207EBXDBXD6683MR1207E.CEL0.0170.0121.681136.8660.40%38.10%1.50%1.450.77GU
81R2551E1BXDBXD6867FR2551E.CEL0.2940.2912.499254.30%44.10%1.60%2.911.55UTM RW
82R2726EBXDBXD6864MR2726E.CEL0.1250.0251.811153.0958.70%39.80%1.50%1.390.78GU
83R2593E1BXDBXD6959FR2593E.CEL0.0270.0381.6712859.20%39.50%1.30%1.470.92UTM RW
84R2727EBXDBXD6965MR2727E.CEL0.010.0081.578143.8660.30%38.30%1.40%1.340.77GU
85R2537E2BXDBXD7059MR2537E2.CEL0.0490.0922.939958.00%40.50%1.60%1.290.75UTM RW
86R975EBXDBXD7064FR975E.CEL0.0280.0241.841137.9758.00%40.50%1.40%1.360.79GU
87R2779EBXDBXD7364FR2779E.CEL0.0120.0381.746121.1159.60%39.00%1.40%1.50.8GU
88R2565E1BXDBXD7561FR2565E.CEL0.1180.1241.7910258.00%40.50%1.50%2.313.47UTM RW
89R1397E-reBXDBXD7558MR1397E-re.CEL0.0320.011.449189.7159.60%39.00%1.40%1.390.82GU
90R2538E1BXDBXD877FR2538E.CEL0.0330.0561.9110261.20%37.30%1.50%1.520.79UTM RW
91R2709EBXDBXD861MR2709E.CEL0.0120.0111.9999.7960.90%37.60%1.50%1.420.76GU
92R2579E1BXDBXD8065FR2579E.CEL0.0130.0262.427259.20%39.40%1.50%1.730.82UTM RW
93R2686EBXDBXD8061MR2686E.CEL0.0460.052.342119.6356.00%42.60%1.50%1.380.79GU
94R2692EBXDBXD8563FR2692E.CEL0.0060.0071.423160.8760.20%38.30%1.40%1.460.79GU
95R2715EBXDBXD8591MR2715E.CEL0.0070.0081.488142.661.20%37.30%1.40%1.50.78GU
96R1405EBXDBXD8658FR1405E.CEL0.0530.0522.351119.3456.40%42.20%1.40%1.640.81GU
97R2540E1BXDBXD8763MR2540E.CEL0.0140.0342.339361.10%37.40%1.40%1.220.81UTM RW
98R2724EBXDBXD8763FR2724E.CEL0.0130.0191.906113.7160.70%37.90%1.40%1.450.79GU
99R2545E1BXDBXD8967MR2546E.CEL0.2660.2571.6710556.20%42.30%1.50%3.69.84UTM RW
100R1433EBXDBXD8963FR1433E.CEL0.0290.0262.241115.8657.70%40.80%1.50%1.410.78GU
101R2569E1BXDBXD967MR2569E.CEL0.2560.2391.758755.10%43.40%1.50%2.823.14UTM RW
102R2708EBXDBXD960FR2708E.CEL0.0240.0451.966126.4657.70%40.70%1.50%1.40.84GU
103R2578E2BXDBXD9061FR2578E2.CEL0.0410.0622.799258.60%39.80%1.60%1.520.77UTM RW
104R859EBXDBXD9072MR859E.CEL0.0280.021.847152.2257.90%40.70%1.40%1.360.77GU
105R2554E1BXDBXD9667MR2554E.CEL0.0050.0082.189360.20%38.30%1.50%1.460.77UTM RW
106R2733EBXDBXD9667FR2733E.CEL0.0240.0541.7113.9962.10%36.60%1.30%1.40.78GU
107R2577E1BXDBXD9755MR2577E.CEL0.0650.0692.077759.50%39.10%1.40%1.871.29UTM RW
108R2649EBXDBXD9774FR2649E.CEL0.0290.0322.343119.0457.50%41.20%1.40%1.530.8GU
109R2688EBXDBXD9867MR2688E.CEL0.0320.031.772145.2458.50%40.00%1.50%1.480.81GU
110R1700E1GDPC3H/HeJ83FR1700E.CEL0.1520.1682.986960.80%37.90%1.40%1.480.78UTM RW
111R1704E1GDPC3H/HeJ83MR1704E.CEL0.1540.1652.588860.10%38.60%1.30%1.380.84UTM RW
112R0872E2GDP BXDC57BL/6J66MR0872E.CEL0.0140.0233.138958.90%39.60%1.50%1.30.79UTM RW
113R2607E1GDP BXDC57BL/6J67FR2605E.CEL0.0080.0182.4311558.60%40.00%1.40%1.310.76UTM RW
114R2564E1GDPCAST/Ei64FR2564E.CEL0.1240.1051.948958.50%39.90%1.60%1.60.77JAX
115R2580E1GDPCAST/Ei64MR2580E.CEL0.1230.1092.099558.20%40.10%1.70%1.40.76JAX
116R2600E1GDP BXDD2B6F172FR2600E.CEL0.0080.022.479558.10%40.20%1.70%1.410.78UTM RW
117R2604E1GDP BXDD2B6F169MR2604E.CEL0.0050.0142.669059.40%39.20%1.50%1.280.79UTM RW
118R2572E1GDP BXDDBA/2J65MR2572E.CEL0.0910.1062.417955.50%42.90%1.60%1.370.79UTM RW
119R2636E1GDPKK/HIJ64FR2636E.CEL0.0440.0432.619358.90%39.50%1.50%1.390.76UTM RW
120R2637E1GDPKK/HIJ64MR2637E.CEL0.0560.0362.1910359.40%39.00%1.50%1.30.79UTM RW
121R0999E1GDPLG/J57FR0999E.CEL0.0210.0232.458259.40%39.10%1.50%1.380.79UTM RW
122R1004E1GDPLG/J65MR1004E.CEL0.0250.0282.449258.70%39.80%1.50%1.380.79UTM RW
123R1688E1GDPNOD/LtJ66FR1688E.CEL0.0280.0332.669858.60%39.90%1.50%1.260.8JAX
124R2566E1GDPNOD/LtJ76MR2566E-2.CEL0.0360.043.036959.80%38.80%1.50%1.380.75UTM RW
125R2535E1GDPNZO/H1LtJ62FR2535E.CEL0.0370.0621.898660.40%38.20%1.40%1.410.85JAX
126R2550E1GDPNZO/HILtJ96MR2550E.CEL0.0250.0291.798760.70%37.80%1.50%1.520.82JAX
127R2634E1GDPPWD/PhJ62FR2635E.CEL0.1260.1143.299055.90%42.50%1.60%1.570.81JAX
128R2635E1GDPPWD/PhJ62MR2634E.CEL0.150.1373.728054.20%44.10%1.70%1.530.85JAX
129R2544E1GDPPWK/PhJ63FR2544E.CEL0.1740.1752.210854.90%43.50%1.70%1.360.82JAX
130R2549E1GDPPWK/PhJ83MR2549E.CEL0.1030.0872.288457.30%41.20%1.50%1.570.83JAX
131R2368E1GDPWSB/EI67FR2368E.CEL0.0410.0472.578659.50%39.10%1.40%1.290.74UTM RW
132R2547E1GDPWSB/Ei67MR2547E.CEL0.0410.0392.149058.20%40.10%1.60%1.320.77UTM RW
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    About downloading this data set:

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This data set is not yet available as a bulk download. Please contact Robert W. Williams to request special data access.

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    About the array platfrom:

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Affymetrix Mouse Genome 430 2.0 arrays: The 430 2.0 array consists of 992936 25-nucleotide probes that estimate the expression of approximately 39,000 transcripts (many are near duplicates). The array sequences were selected late in 2002 using Unigene Build 107. The array nominally contains the same probe sequences as the old M430A and 430B array pair. However, we have found that roughy 75000 probes differ between those on A and B arrays and those on the new 430 2.0.

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    About data processing:

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Probe (cell) level data from the CEL file: These CEL values produced by GCOS are 75% quantiles from a set of 91 pixel values per cell. The CEL files were processed using the RMA protocol. We processed the three batches together in RMA. + + +
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  • Step 1: We added an offset of 1.0 unit to each cell signal to ensure that all values could be logged without generating negative values. We then computed the log base 2 of each cell. + +
  • Step 2: We performed a quantile normalization of the log base 2 values for the total set of arrays using the same initial steps used by the RMA transform. + +
  • Step 3: We computed the Z scores for each cell value. + +
  • Step 4: We multiplied all Z scores by 2. + +
  • Step 5: We added 8 to the value of all Z scores. The consequence of this simple set of transformations is to produce a set of Z scores that have a mean of 8, a variance of 4, and a standard deviation of 2. The advantage of this modified Z score is that a two-fold difference in expression level corresponds approximately to a 1 unit difference. + +
  • Step 7: Finally, when appropriate, we computed the arithmetic mean of the values for the set of microarrays for each strain. Technical replicates were averaged before computing the mean for independent biological samples. + +
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After RMA processing all arrays were rank-order normalized. This second round of quantile normalization removes much residual non-linearity across arrays and forces every array to have the same distribution of values as the mean of all arrays. Comparative array data quality was then evaluated in DataDesk. Outlier arrays were flagged by visual inspection in DataDesk, usually by means of an analysis of scatter plots and more quantitatively by generating a correlation matrix of all arrays. Those arrays with mean correlation <0.96 versus all other arrays indicates trouble or a biological outlier). In some cases, outliers were expected, such as samples from strains with retinal degeneration (FVB/NJ, NOD/LtJ, MOLF/EiJ, C3H/HeJ and BXD24) and samples from wild subspecies such as WSB/EiJ, CAST/Ei, PWD/Ph, and PWK/Ph. However, when arrays were anomolous both within strain and across strains, they were often simply discarded. We tended to keep arrays that "conformed" to the expectation. The assumption in these cases is that anomolous data are much more likely due to experimental problems and errors than to informative biological variation. Approximately 11 (CHECK) arrays total were discarded in batches 1, 2, and 3 combined. + +

After this process, the acceptable set of arrays was renormalized using all step as above, starting with the original RMA procedure, etc. + + + +

We reviewed the data set using a new method developed by RW Williams, Jeremy Peirce, and Hongqiang Li. For the full set of arrays that passed standard QC protocols described above, we computed the strain means for the BXD strains, B6, D2, and F1s. Using this set of strain means we then computed LRS scores for all 45101 probe sets and counted the number of transcripts that generated QTLs with LRS values greater than 50. This value (e.g. 1800) represented the QTL harvest for the full data set. We then dropped a single array from the data set, recomputed strain means, and recomputed the number of transcripts with LRS scores great than 50. This value is expected to typically reduce the number of QTLs that reach the criterion level (e.g., 1750 QTLs > 50). This process was repeated for every array to obtain an array-specific difference value--the effect of removing that array on the total QTL count. For example, the loss of a single array might cause a decrease in 50 QTLs. Values ranged from approximately -90 (good arrays) to +40 (bad arrays). This procedure is similar in some ways to a jackknife protocol, although we are not using this procedure to esimate an error term, but rather as a method to polish a data set. + +

During this final process we discovered that nearly XX arrays in the second batch had been mislabeled at some point in processing. We computed the correct strain membership of each array using a large number of Mendelian probe sets (more than 50) and comparing their match to standard SNP and microsatellite markers and the original array data set of November 2005. This allowed us to rescue a large number of arrays that were of very high quality. + +

A third batch of approximately 40 arrays were processed by Yan Jiao and Wiekuan Gu in August 2006. These complete data set assembled by Hongqiang Li. This process again included a correction for a batch effect. + +

For this June 2006 data set Hongqiang Li used a new batch correction method that stabilizes the range of expression in each batch. For each of the three large batches, we extracted the minumum and maximum raw probe expression (CEL file level) value. We then adjusted raw probe values in each batch to have the same range as the largest batch (Batch 1) using a simple linear interpolation. These procedure generated new correct CEL files which were then used with RMA. (note added by RWW and HQL, Oct 19, 2006) + + + +

    Data source acknowledgment:

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+Support for acquisition of microarray data sets was generously provided by Dr. Barrrett Haik, Chair of the Department of Ophthalmology, and director of the Hamilton Eye Institute. Support for the continued development of GeneNetwork was provided by a NIDA/NIMH/NIAAA Human Brain Project grant. All arrays were processed at the VA Medical Center, Memphis by Yan Jiao and Weikuan Gu. +
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    Information about this text file:

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This text file originally generated by RWW, May 26, 2006. Updated by RWW, Oct 10, 2006. +

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