|
| Status |
Public on Feb 17, 2012 |
| Title |
adult_bone_marrow_proB |
| Sample type |
RNA |
| |
|
| Source name |
adult bone marrow pro-B cells
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57BL/6
developmental stage: adult
tissue: bone marrow
cell type: B220+CD43+IgM-veCD19+CD24+ pro-B cells
|
| Biomaterial provider |
Joan Yuan
|
| Treatment protocol |
FACS-sorted pro-B cells (B220+CD43+IgM-veCD19+CD24+) from the bone marrow of 3 adult mice were pooled.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNAzol (Molecular Research Center, Inc) was used per manufacturer's instructions to extract total RNA. 100 ng total RNA was used as starting material. RNA concentration was determined using the Qubit RNA broad range assay and fluorometer (Invitrogen). To confirm the concentration and determine the integrity of the RNA, the Eukaryote Total RNA nano Series II chip on a 2100 Bioanalyzer (Agilent) was used.
|
| Label |
NanoString oligonucleotide tag
|
| Label protocol |
Sample labelling was performed per manufacturer’s instructions at the DNA Sequencing and Digital Expression Core Facility (National Cancer Institute).
|
| |
|
| Hybridization protocol |
Flow-cell hybridization was performed per manufacturer’s instructions at the DNA Sequencing and Digital Expression Core Facility (National Cancer Institute).
|
| Scan protocol |
Scanning was performed per manufacturer’s instructions at the DNA Sequencing and Digital Expression Core Facility (National Cancer Institute).
|
| Data processing |
Excel was used for data normalization. Per manufacturer's instructions, data were normalized to the sum of positive control count values provided in assay to account for lane to lane variation. Then the data were normalized again to the sum of the counts of the 75 highest expressing miRNAs in each sample. let-7 miRNAs that fell into the category of the 75 highest expressing miRNAs were excluded from the list. Thus, let-7a, let-7b, let-7c, let-7d, let-7f, let-7g and let-7i were excluded for adult pro-B. Background levels were determined by calculating the mean plus two standard deviations of eight negative control detectors. By this method, a normalized count ≤40 is considered undetectable for this dataset.
|
| |
|
| Submission date |
Jan 14, 2012 |
| Last update date |
Feb 17, 2012 |
| Contact name |
Stefan Adi Muljo |
| E-mail(s) |
[email protected]
|
| Phone |
301-594-2116
|
| Fax |
301-480-7929
|
| URL |
http://www3.niaid.nih.gov/labs/aboutlabs/li/iiu/
|
| Organization name |
National Institutes of Health
|
| Department |
National Institute of Allergy and Infectious Diseases
|
| Lab |
Laboratory of Immunology
|
| Street address |
10 Center Drive
|
| City |
Bethesda |
| State/province |
MD |
| ZIP/Postal code |
20982-1892 |
| Country |
USA |
| |
|
| Platform ID |
GPL15089 |
| Series (2) |
| GSE35107 |
microRNA profiling of fetal liver and adult bone marrow pro-B cells |
| GSE35214 |
Lin28b expression distinguishes fetal versus adult lymphopoiesis |
|
