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Sample GSM6924631 Query DataSets for GSM6924631
Status Public on Jun 13, 2024
Title C32
Sample type RNA
Source name Peripheral blood
Organism Homo sapiens
Characteristics gender: Male
tissue: Blood
disease state: healthy
Extracted molecule total RNA
Extraction protocol Ribopure Blood RNA Isolation Kit by Thermo Fisher was utilized to extract the RNA (including small RNAs) as suggested protocol by manufacturer protocol.
Label protocol A TaqMan Custom RT pool primer (Cat#A25630) of 112 miRNAs was used for cDNA synthesis by reverse transcription. Reverse transcription of mature miRNAs was carried out by using a TaqMan miRNA Reverse Transcription kit reagents (Cat# 4366597) from Thermo Fischer, USA. In short, the reaction mixture contained MultiScribe reverse transcriptase 75 units, RT buffer 1X, OpenArray Custom RT primers 1X, RNase inhibitor 2 units, 3mM MgCl2, 2.0mM deoxynucleotide triphosphates (dNTPs) and 100ng of RNA input limiting 7.5µl the final reaction volume. Thermal cycling of Reverse transcription process was repeated for additional 39 cycles of 16°C for 2 minutes, 42°C for 1 minute and 50°C for 1 second. . In Preamplification, a total 25 µl reaction volume including 2.5µl custom preamp primer pools mixed with 12.5µl of preamp master mix and 2.5µl cDNA and additional nuclease-free water add to reach the final volume. The thermal cycling condition used for preamplification were 95°C for 10 min, 55°C for 2 min, 72°C for 2 min and 12 cycles of 95°C/15 sec, 60°C/4 min. At the end of cycling, preamplification products were incubated at 99.90C for 10 min. for inactivation of enzymes. Preamplification products were diluted (1 to 40) in 0.1X Tris-EDTA buffer. The diluted preamplification products mixed with 2x TaqMan OpenArray Real-Time PCR Master Mix (Cat#4462159) and pipette on OpenArray plates by OpenArray AccuFill System from Thermo Fisher Scientific and run the preamplification product as described manufacturer on QuantStudio 12K Flex RT-PCR machine from Thermo Fisher Scientific, USA
Hybridization protocol N/A
Scan protocol N/A
Description Raw data file:
Control 32
Data processing The raw data was analyzed by ExpressionSuite software and Relative quantification of miRNAs expression was calculated by -ΔΔCt method, selecting U6 snRNA level as endogenous control. software performs all deltadeltaCt based fold-change calculations and performed student t-test to calculate p-value from the uploaded raw threshold cycle data .
Normalized values and fold-change data are available on the series record.
Submission date Jan 09, 2023
Last update date Jun 13, 2024
Contact name SANJAY YADAV
Organization name AIIMS RAEBARLI
Street address All India Institute of Medical Sciences Dalmau Road, Munshiganj,
City Raebareli
State/province UTTAR PRADESH
ZIP/Postal code 229405
Country India
Platform ID GPL33008
Series (1)
GSE222480 Transcriptomics and proteomics approach for the identification of altered blood microRNAs and plasma proteins in Parkinson’s disease by Custom Brain Specific miRNA OpenArray Real-time PCR panel

Data table header descriptions
VALUE non-normalized data

Data table
1 16.416
2 20.804
3 19.283
4 22.414
5 28.069
6 30.239
8 23.678
10 26.853
11 24.766
13 23.768
16 12.562

Total number of rows: 112

Table truncated, full table size <1 Kbytes.

Supplementary data files not provided
Processed data are available on Series record

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