MARC details
| 000 -LEADER |
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| fixed length control field |
02696nam a2200241Ia 4500 |
| 003 - CONTROL NUMBER IDENTIFIER |
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| control field |
MX-MdCICY |
| 005 - DATE AND TIME OF LATEST TRANSACTION |
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| control field |
20250625153940.0 |
| 040 ## - CATALOGING SOURCE |
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| Transcribing agency |
CICY |
| 090 ## - LOCALLY ASSIGNED LC-TYPE CALL NUMBER (OCLC); LOCAL CALL NUMBER (RLIN) |
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| Classification number (OCLC) (R) ; Classification number, CALL (RLIN) (NR) |
B-14402 |
| 008 - FIXED-LENGTH DATA ELEMENTS--GENERAL INFORMATION |
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| fixed length control field |
250602s9999 xx |||||s2 |||| ||und|d |
| 245 10 - TITLE STATEMENT |
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| Title |
Effect of experimental treatment on housekeeping gene expression: validation by real-time, quantitative RT-PCR |
| 490 0# - SERIES STATEMENT |
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| Volume/sequential designation |
J. Biochem. Biophys. Methods, 46(1-2), p.69-81, 2000 |
| 520 3# - SUMMARY, ETC. |
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| Summary, etc. |
The effects of serum on the expression of four commonly used housekeeping genes were examined in serum-stimulated fibroblasts in order to validate the internal control genes for a quantitative RT-PCR assay. NIH 3T3 fibroblasts transfected with an inducible chimeric gene were serum-starved for 24 h and then induced with 15 percent serum for 8 h. Serum did not alter the amount of total RNA that was expressed in the cells, however, the amount of mRNA significantly increased over time with serum-stimulation. Both messenger and total RNA from each of the time points were reverse transcribed under two different conditions; one in which the reactions were normalized to contain equal amounts of RNA and another series of reactions that were not normalized to RNA content. The resulting cDNA was amplified by real-time, quantitative PCR using gene-specific primers for b-actin, b-2 microglobulin, glyceraldehyde-3-phosphate dehydrogenase (GAPDH)and 18S ribosomal RNA. The expression of b-actin and GAPDH increased up to nine- and three-fold, respectively, under all conditions of reverse transcription (P,0.01). The expression of 18S rRNA increased with serum-stimulation when the cDNA synthesized from non-normalized, total RNA was assayed (P,0.01)but not when the reverse transcriptions were normalized to RNA content (P.0.05). The expression of b-2 microglobulin increased up to two-fold when assayed from cDNA synthesized from non-normalized mRNA, but was unaffected by serum when the reverse transcriptions were normalized to mRNA. b-2 Microglobulin non-normalized reverse transcription reactions. Thus, b-2 microglobulin and 18S rRNA are suitable internal control genes in quantitative serum-stimulation studies, while b-actin and GAPDH are not. The internal control gene needs to be properly validated when designing quantitative gene expression studies. |
| 650 14 - SUBJECT ADDED ENTRY--TOPICAL TERM |
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| Topical term or geographic name entry element |
REAL-TIME PCR |
| 650 14 - SUBJECT ADDED ENTRY--TOPICAL TERM |
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| Topical term or geographic name entry element |
QUANTITATIVE RT-PCR |
| 650 14 - SUBJECT ADDED ENTRY--TOPICAL TERM |
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| Topical term or geographic name entry element |
GENE EXPRESSION |
| 650 14 - SUBJECT ADDED ENTRY--TOPICAL TERM |
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| Topical term or geographic name entry element |
SYBR GREEN |
| 650 14 - SUBJECT ADDED ENTRY--TOPICAL TERM |
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| Topical term or geographic name entry element |
SERUM-STIMULATION |
| 700 12 - ADDED ENTRY--PERSONAL NAME |
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| Personal name |
Schmittgen, T.D. |
| 700 12 - ADDED ENTRY--PERSONAL NAME |
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| Personal name |
Zakrajsek, B.A. |
| 856 40 - ELECTRONIC LOCATION AND ACCESS |
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| Uniform Resource Identifier |
<a href="https://drive.google.com/file/d/10tetQgkfU8yb2xjJL6kH3CSrjLKgPCpu/view?usp=drivesdk">https://drive.google.com/file/d/10tetQgkfU8yb2xjJL6kH3CSrjLKgPCpu/view?usp=drivesdk</a> |
| Public note |
Para ver el documento ingresa a Google con tu cuenta: @cicy.edu.mx |
| 942 ## - ADDED ENTRY ELEMENTS (KOHA) |
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| Source of classification or shelving scheme |
Clasificación local |
| Koha item type |
Documentos solicitados |
