|
| Status |
Public on Mar 17, 2011 |
| Title |
Footpads_M. leprae infected_12 month_rep1 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
M. lepare non-infected footpads
|
| Organism |
Mus musculus |
| Characteristics |
tissue: footpads
strain: CAnN.Cg-Foxn1nu/CrljBgi
experimental period: 12 months
|
| Treatment protocol |
M. leprae non-infected footpads: PBS (subcutaneous injection), M. leprae-infected footpads: 2´10^6 bacteria (subcutaneous injection)
|
| Extracted molecule |
total RNA |
| Extraction protocol |
(1) Total RNA extracted using Trizol following manufacturer's instructions, (2) DNA removal using Rneasy RNA extraction column following manufacturer's instructions
|
| Label |
Cy3
|
| Label protocol |
Direct Labeling (20 µg of total RNA were primed with oligo dT(18) primer at 70°C for 10 min, then reversed transcribed at 42°C for 1 h in the presence of Reverse transcriptase, and dNTP, cyanine dCTP (control - Cy3 and sample-Cy5)
Labeled cDNA were purifed with 3M sodium acetate and absoulte ethanol, checked quality of synthesized labeled-cDNA (Spectrophotometer A260, A650, A550), and the calculated concentration and incorporation rate
|
| |
|
| Channel 2 |
| Source name |
M. lepare-infected footpads
|
| Organism |
Mus musculus |
| Characteristics |
tissue: footpads
strain: CAnN.Cg-Foxn1nu/CrljBgi
period: 12 months
|
| Treatment protocol |
M. leprae non-infected footpads: PBS (subcutaneous injection), M. leprae-infected footpads: 2´10^6 bacteria (subcutaneous injection)
|
| Extracted molecule |
total RNA |
| Extraction protocol |
(1) Total RNA extracted using Trizol following manufacturer's instructions, (2) DNA removal using Rneasy RNA extraction column following manufacturer's instructions
|
| Label |
Cy5
|
| Label protocol |
Direct Labeling (20 µg of total RNA were primed with oligo dT(18) primer at 70°C for 10 min, then reversed transcribed at 42°C for 1 h in the presence of Reverse transcriptase, and dNTP, cyanine dCTP (control - Cy3 and sample-Cy5)
Labeled cDNA were purifed with 3M sodium acetate and absoulte ethanol, checked quality of synthesized labeled-cDNA (Spectrophotometer A260, A650, A550), and the calculated concentration and incorporation rate
|
| |
|
| |
| Hybridization protocol |
Purifed cDNA were denatured, and the samples were applied to microarrays enclosed in hybridization chambers. After hybridization, slides were washed sequential ((1) 2XSSC, 0.1%SDS, (2) 1XSSC, (3) 0.2XSSC) and were dried by centrifuge
|
| Scan protocol |
Scanned on an Genepix 4000B scanner.
Images were quantified using GenePix pro Software (version 5.1).
|
| Description |
Footpads_M. leprae infected_12 month_rep1
|
| Data processing |
Box plot and Intesity dependent (LOWESS) normalized, background subtracted data obtained from log2 of processed Red signal/processed Green signal. GenePix pro 5.1 was used.
|
| |
|
| Submission date |
Jul 07, 2010 |
| Last update date |
Mar 17, 2011 |
| Contact name |
Tae Hoon Kim |
| E-mail(s) |
ecotype531@catholic.ac.kr
|
| Phone |
82-2-2258-7451
|
| Fax |
82-2595-2241
|
| URL |
http://www.hansen.re.kr
|
| Organization name |
Medical College of the Catholic University
|
| Department |
Pathology
|
| Lab |
Institute of Hansen's Disease
|
| Street address |
505 Bonpo-dong
|
| City |
Seoul |
| State/province |
Socho-gu |
| ZIP/Postal code |
138-701 |
| Country |
South Korea |
| |
|
| Platform ID |
GPL10650 |
| Series (1) |
| GSE22784 |
Profiling of Gene expression in Mycobacterium leprae (M. leprae)-infected footpad of BALB/c nu/nu mice |
|
