|
| Status |
Public on Apr 28, 2022 |
| Title |
Yak 3 +120d mammary gland tissue |
| Sample type |
RNA |
| |
|
| Source name |
Yak 3 mammary gland tissue
|
| Organism |
Bos grunniens |
| Characteristics |
tissue: mammary gland
gender: female
days relative to parturition: +120days
|
| Treatment protocol |
Biopsy of the mammary gland was performed by a veterinarian. Approximately 1 g of tissue was obtained alternatively at each time point from the left and right rear quarter on the mammary gland. Briefly, to obtain mammary parenchyma, the mammary capsule was incised and blunt dissection was performed. Fat and connective tissue were dissected, and clean mammary parenchyma subsequently washed with diethyl pyro carbonate-treated water. Tissue was cut into small pieces and immediately stored in liquid nitrogen for total RNA extraction.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA containing small RNA was extracted from yak mammry gland tissue by using the Trizol reagent (Invitrogen) and purified with mirVana miRNA Isolation Kit (Ambion, Austin, TX, USA) according to manufacter’s protocol.
|
| Label |
Cy3
|
| Label protocol |
cDNA labeled with a fluorescent dye (Cy3-dCTP) was produced by Eberwine’s linear RNA amplification method with subsequent enzymatic reaction. In detail, double-stranded cDNA (containing the T7 RNA polymerase promoter sequence) was synthesized from 100 ng total RNA using the CbcScript reverse transcriptase with cDNA synthesis system according to the manufacturer’s protocol (Capitalbio) with the T7 Oligo (dT). Germany). The Klenow enzyme labeling strategy was adopted after reverse transcription using CbcScript Ⅱ reverse transcriptase. Labeled cDNA was purified with a PCR NucleoSpin Extract II Kit (Macherey-Nagel, Düren, Germany) and resuspended in elution buffer.
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| |
|
| Hybridization protocol |
DNA in hybridization solution was denatured at 95℃ for 3 min prior to loading onto a microarray.Array hybridization was performed in an Agilent Hybridization Oven overnight at a rotation speed of 20 rpm at 42℃ and washed with two consecutive solutions (0.2% SDS, 2× SSC at 42℃ for 5 min, and 0.2× SSC for 5 min at room temperature).
|
| Scan protocol |
The cleaned chip was scanned by Agilent chip scanner (g2565ca) to obtain the hybridization image.
|
| Description |
Gene expression of Yak mammary gland at 9 time points ralatead to parturition
|
| Data processing |
Agilent feature extraction (v10.7) software was used to analyze the hybrid images and extract the data. Thwn genespring software V12 (Agilent) for data analysis.
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| |
|
| Submission date |
Apr 25, 2022 |
| Last update date |
Apr 28, 2022 |
| Contact name |
Yu Wang |
| Organization name |
Southwest Minzu University
|
| Street address |
#16, South Section, 1st Ring Road
|
| City |
Chengdu |
| State/province |
Sichuan |
| ZIP/Postal code |
610041 |
| Country |
China |
| |
|
| Platform ID |
GPL11649 |
| Series (1) |
| GSE201438 |
Dynamic profiles of the Yak mammary transcriptome during lactation |
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