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Status |
Public on Apr 30, 2007 |
Title |
Fruit6_woolly_rep4 |
Sample type |
RNA |
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Source name |
Mesocarp tissue of O´Henry cultivar peach
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Organism |
Prunus persica |
Characteristics |
Woolly fruit (0% juice)
|
Biomaterial provider |
Chilean Functional Genomics in Prunus persica Consortium: Universidad de Chile, Instituto de Investigaciones Agropecuarias, Asociacion de Exportadores, Fundacion para el Desarrollo Fruticola and Fundacion Chile. Supported by FDI-CORFO (FDI G02 P1001)
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Treatment protocol |
Peach sampled from commercial harvest and exposed 7 days at 21 ºC after 15 days at 4ºC
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Extracted molecule |
polyA RNA |
Extraction protocol |
mRNA was purified using an Oligotex Mini kit (Qiagen), following manufacturer recommendations
|
Label |
32P
|
Label protocol |
The 32P-labeled target DNA samples were prepared from poly(A)+ RNAs by incorporation of [α-32P]dCTP during first-strand cDNA synthesis, according to the protocol described by Bernard et al. (1996), unincorporated radioactive nucleotides were removed using the QIAquick Nucleotide Removal according to the manufacturer’s instructions (Qiagen).
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Hybridization protocol |
Pre-hybridization in 5 ml of hybridization buffer (5X SSC, 5 X Denhardt solution, 1% SDS, and 50% formamide) for 1-3 h at 42 ºC. Hybridization was performed in the same buffer for 16-18 h at 42 ºC.
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Scan protocol |
After washing, the membranes were sealed with a plastic film and exposed for 72-96 h to Phosphor screens (Kodak).
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Description |
Woolly fruit (0% juice)
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Data processing |
Quantification of the signal intensity was performed using the VersArray Analyzer software (BioRad). Raw values were measured as the volume of pixels within a circle encompassing the spot. Local background values were measured in the corners of spot and were subtracted from the signal intensity values for each spot. Spots that showed: (1) Signal Mean < (Background Mean + (1 x Background Standard Deviation; and (2) qcom < 0.8, were considered as low-quality spots and were removed. Qcom values were calculated as described in Wang et al., 2001. Then, the net intensity value of each spot was normalized by that of dap, a spike mRNA control.
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Submission date |
Feb 27, 2007 |
Last update date |
Mar 02, 2007 |
Contact name |
Mauricio Gonzalez |
E-mail(s) |
maugonzalez@inia.cl
|
Organization name |
Instituto de Investigaciones Agropecuarias
|
Lab |
Postcosecha
|
Street address |
Santa Rosa 11610
|
City |
Santiago |
State/province |
Region Metropolitana |
ZIP/Postal code |
439-3 |
Country |
Chile |
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Platform ID |
GPL4889 |
Series (1) |
GSE7145 |
Transcriptional changes associated with woolliness during fruit ripening in Prunus persica |
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