|
| Status |
Public on Dec 30, 2012 |
| Title |
Hearts_Normal_Control |
| Sample type |
RNA |
| |
|
| Source name |
3 pooled normal hearts of BALB/c mice
|
| Organism |
Mus musculus |
| Characteristics |
donor: n/a
strain: Balb/c
rejection: n/a
|
| Growth protocol |
Eight to ten-week-old C57BL/6 and BALB/c mice were used. Syngeneic (C57BL/6 to C57BL/6) and allogeneic (BALB/c to C57BL/6) heterotopic heart transplants were performed.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated from heart grafts, GILs and PBLs using the mirVana miRNA isolation kit (Applied Biosystems, Foster City, CA). Total RNA isolated from hearts of normal BALB/c mice (donor) and PBLs of normal C57BL/6 (recipient) mice were used as controls and calibrators for microarray data analysis. Three individual RNA samples from either transplanted or normal animals were pooled together (100 ng of each, 300 ng in total) for cDNA preparation using TaqMan microRNA reverse transcription kit and the corresponding Megaplex primer pools (Applied Biosystems). The resulting cDNA samples underwent preamplification using TaqMan PreAmp master mix and corresponding Megaplex PreAmp primers (Applied Biosystems).
|
| Label |
FAM
|
| Label protocol |
The PCR reaction was carried out on an Applied BioSystems 7900HT thermocycler.
|
| |
|
| Hybridization protocol |
n/a
|
| Scan protocol |
n/a
|
| Description |
Normal hearts
|
| Data processing |
Data was analyzed using SDS and RQ software (Applied Biosystems). Fold changes for each miRNA were normalized to the endogenous control snoRNA202. The relative expression levels were calculated using the 2-∆∆Ct method. For heart grafts, normal BALB/c hearts were used as a calibrator. For GILs, normal C57BL/6 PBMC was used as a calibrator.
|
| |
|
| Submission date |
Oct 02, 2011 |
| Last update date |
Dec 30, 2012 |
| Contact name |
Liang Wei |
| E-mail(s) |
liangwei@stanford.edu
|
| Phone |
+1 650-725-6616
|
| Fax |
+1 650-498-6250
|
| URL |
http://www.stanford.edu/group/TIL/
|
| Organization name |
Stanford University
|
| Department |
Surgery
|
| Lab |
Transplant Immunobiology Laboratory
|
| Street address |
1201 Welch Rd MSLS P328
|
| City |
Stanford |
| State/province |
CA |
| ZIP/Postal code |
94305 |
| Country |
USA |
| |
|
| Platform ID |
GPL14646 |
| Series (1) |
| GSE32545 |
MicroRNAs as Regulators of the Immune Response during Allograft Rejection |
|
