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| Status |
Public on Oct 08, 2000 |
| Title |
Dmel.testis.1.0 |
| Technology type |
spotted DNA/cDNA |
| Distribution |
non-commercial |
| Organism |
Drosophila melanogaster |
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| Description |
The array was assembled using 1,681 testes ESTs, representing 1,527 Drosophila melanogaster EST sequences and 144 anonymous clones from the bs collection (CLONE_ID).
An aliquot of the same DNA preparation used in the sequencing reactions (GB_ACC), diluted 2/50 in TE, provided material for the PCRs.
Inserts were PCR amplified (1X PCR buffer [Life Technologies], 1.5 mM MgCl2, 0.2 mM dNTPs, 1 µM T7 22-mer primer, 1 µM T3 20-mer primer, 2 µl of plasmid DNA dilution [approximately 10 ng], 0.035 U/µl recombinant Taq DNA polymerase [Life Technologies], in a volume of 100 µl, cycled 94C 2 min, 30X[94 C 0.5 min, 55 C 0.5 min, 2.5 min 72 C], 10 min 72 C in a PTC-225 DNA Engine Tetrad [MJ Research]), and successful amplification was confirmed by agarose gel electrophoresis (PCR-QC: passed = product present, failed = no product).
Samples of approximately 5 nl of 250 µg/ml DNA, in 0.1 N NaOH were printed in 300 µm spots in subarrays (SUB-ARRAY) of 12 x 12 clones (ROW and COLUMN indicated) at 665 µm spacing on Supercharge nytran membranes (Schleicher & Schuell) using a GMS 417 arrayer (Genetic Microsystems).
Subarrays were printed in duplicate (DUPLICATE a or b).
Printing was validated (SPOT_QC) by hybridization of one array with the short vector sequence common to all amplicons (pBluescript SK, 626-791 bp).
SPOT_QC values are hybridization intensity. BEST-HIT and associated E value for ESTs aligned with the nr database are given.
Keywords = germline sex testis, membrane, filter
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| Contributor(s) |
Oliver B, Andrews J, Bouffard GG, Cheadle C, Lu J, Becker KG |
| Citation(s) |
11116097 |
| Submission date |
Oct 03, 2000 |
| Last update date |
Oct 28, 2005 |
| Contact name |
Brian Oliver |
| E-mail(s) |
[email protected]
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| Phone |
301-204-9463
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| Organization name |
NIDDK, NIH
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| Department |
LBG
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| Lab |
Developmental Genomics
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| Street address |
50 South Drive
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| City |
Bethesda |
| State/province |
MD |
| ZIP/Postal code |
20892 |
| Country |
USA |
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| Samples (8)
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GSM3, GSM4, GSM5, GSM6, GSM7, GSM8
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| Series (1) |
| GSE462 |
Analysis of transcription in the Drosophila melanogaster testis |
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| Data table header descriptions |
| ID |
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| GB_ACC |
GenBank accession number |
| BSCC_ID |
bs collection clone identifier |
| CLONE_ID |
Flybase identifier |
| SUB-ARRAY |
sub-array number |
| DUPLICATE |
dupilcate number |
| ROW |
row number |
| COLUMN |
column number |
| PCR_QC |
PCR quality control |
| SPOT_ID |
spot identifier |
| BEST_HIT |
BLAST best hit against nr database |
| E_VAL |
E value of best hit |
| SPOT_QC |
spot quality control |
Total number of rows: 3456
| Supplementary data files not provided |
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