PMID- 26872627
OWN - NLM
STAT- MEDLINE
DCOM- 20161108
LR  - 20210109
IS  - 1873-2534 (Electronic)
IS  - 0165-2427 (Linking)
VI  - 170
DP  - 2016 Feb
TI  - Identification of stable reference genes for quantitative PCR in cells derived 
      from chicken lymphoid organs.
PG  - 20-4
LID - S0165-2427(16)30001-0 [pii]
LID - 10.1016/j.vetimm.2016.01.001 [doi]
AB  - Quantitative polymerase chain reaction (qPCR) is a powerful technique for 
      quantification of gene expression, especially genes involved in immune responses. 
      Although qPCR is a very efficient and sensitive tool, variations in the enzymatic 
      efficiency, quality of RNA and the presence of inhibitors can lead to errors. 
      Therefore, qPCR needs to be normalised to obtain reliable results and allow 
      comparison. The most common approach is to use reference genes as internal 
      controls in qPCR analyses. In this study, expression of seven genes, including 
      beta-actin (ACTB), beta-2-microglobulin (B2M), glyceraldehyde-3-phosphate dehydrogenase 
      (GAPDH), beta-glucuronidase (GUSB), TATA box binding protein (TBP), alpha-tubulin 
      (TUBAT) and 28S ribosomal RNA (r28S), was determined in cells isolated from 
      chicken lymphoid tissues and stimulated with three different mitogens. The 
      stability of the genes was measured using geNorm, NormFinder and BestKeeper 
      software. The results from both geNorm and NormFinder were that the three most 
      stably expressed genes in this panel were TBP, GAPDH and r28S. BestKeeper did not 
      generate clear answers because of the highly heterogeneous sample set. Based on 
      these data we will include TBP in future qPCR normalisation. The study shows the 
      importance of appropriate reference gene normalisation in other tissues before 
      qPCR analysis.
CI  - Copyright (c) 2016 Elsevier B.V. All rights reserved.
FAU - Borowska, D
AU  - Borowska D
AD  - The Roslin Institute and R(D)SVS, University of Edinburgh, Easter Bush, 
      Midlothian EH25 9RG, United Kingdom. Electronic address: 
      dominika.borowska@roslin.ed.ac.uk.
FAU - Rothwell, L
AU  - Rothwell L
AD  - The Roslin Institute and R(D)SVS, University of Edinburgh, Easter Bush, 
      Midlothian EH25 9RG, United Kingdom.
FAU - Bailey, R A
AU  - Bailey RA
AD  - Aviagen Ltd., Edinburgh EH28 8SZ, United Kingdom.
FAU - Watson, K
AU  - Watson K
AD  - Aviagen Ltd., Edinburgh EH28 8SZ, United Kingdom.
FAU - Kaiser, P
AU  - Kaiser P
AD  - The Roslin Institute and R(D)SVS, University of Edinburgh, Easter Bush, 
      Midlothian EH25 9RG, United Kingdom.
LA  - eng
GR  - BBS/E/D/20320000/BB_/Biotechnology and Biological Sciences Research 
      Council/United Kingdom
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20160106
PL  - Netherlands
TA  - Vet Immunol Immunopathol
JT  - Veterinary immunology and immunopathology
JID - 8002006
SB  - IM
MH  - Animals
MH  - *Chickens
MH  - Gene Expression Profiling/methods
MH  - *Genes
MH  - Lymphoid Tissue/*cytology/metabolism
MH  - Real-Time Polymerase Chain Reaction/*standards
MH  - Reference Standards
OTO - NOTNLM
OT  - Chicken
OT  - Lymphoid tissues
OT  - Normalisation
OT  - Reference gene
OT  - qPCR
EDAT- 2016/02/14 06:00
MHDA- 2016/11/09 06:00
CRDT- 2016/02/14 06:00
PHST- 2015/07/29 00:00 [received]
PHST- 2015/12/21 00:00 [revised]
PHST- 2016/01/05 00:00 [accepted]
PHST- 2016/02/14 06:00 [entrez]
PHST- 2016/02/14 06:00 [pubmed]
PHST- 2016/11/09 06:00 [medline]
AID - S0165-2427(16)30001-0 [pii]
AID - 10.1016/j.vetimm.2016.01.001 [doi]
PST - ppublish
SO  - Vet Immunol Immunopathol. 2016 Feb;170:20-4. doi: 10.1016/j.vetimm.2016.01.001. 
      Epub 2016 Jan 6.