Comparative Quantitative Study of Surfactant Protein C mRNA by Filter Hybridization and Solution Hybridization in Rats.
10.4046/trd.2001.51.6.517
- Author:
Jin Ho KIM
;
Jang Won SOHN
;
Seok Chul YANG
;
Ho Joo YOON
;
Dong Ho SHIN
;
Sung Soo PARK
- Publication Type:Original Article
- Keywords:
Surfactant protein C mRNA;
Solution hybridization;
Filter hybridization
- MeSH:
Animals;
Base Sequence;
Clone Cells;
Protein C*;
Rats*;
RNA;
RNA, Messenger*
- From:Tuberculosis and Respiratory Diseases
2001;51(6):517-529
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
BACKGROUND: Surfactant protein C(SP-C) is a hydrophobic 5,000 dalton molecule. SP-C has the primary roles in accelerating surface spreading of a surfactant phospholipid. The filter hypbridization and solution hybridization assays are both rapid and sensitive and can be used to measure the RNAs complementary to any cloned DNA sequence. METHODS: The authors measured the SP-C mRNA levels quantitatively using solution hybridization and filter hybridization assays to obtain a standard curve equation to quantify the mRNA of unknown samples comparatively. RESULTS: 1. The minimum level of the specimens by solution hybridization was 3 pg for SP-C mRNA. 2. The standard curve equation of the solution hybridization assay between the counts per minute(Y) and the SP-C mRNA transcript input(X) was Y=6.46 X+244. The correlation coefficient was 0.9. 3. The minimum detection level of specimens by filter hybridization was 0.1 ng for SP-C mRNA. 4. The standard curve equation of the filter hybridization assay between the counts per minute(Y) and SP-C mRNA transcript input(X) is Y=2541.6 X+252.7. The correlation coefficient was 0.99. CONCLUSION: A comparison of CPM/filter in the linear range allowed an accurate and reproducible estimation of the SP-C mRNA copy number. Filter hybridization and solution hybridization assays are both rapid and sensitive and can be used to measure the RNAs complementary to any cloned DNA sequence. It is ideally suited to situations where accurate quantitation of multiple samples is required.
