Abstract：
BACKGROUND:When estimating relative transcript abundances by quantitative real-time PCR (Q-PCR) we found that the results can vary dramatically depending on the method chosen for data analysis. RESULTS:Analyses of Q-PCR results from a salmon louse starvation experiment show that, even with apparently good raw data, different analytical approaches 12 may lead to opposing biological conclusions. CONCLUSION:The results emphasise the importance of being cautious when analysing Q-PCR data and indicate that uncritical routine application of an analytical method will eventually result in incorrect conclusions. We do not know the extent of, or have a universal solution to this problem. However, we strongly recommend caution when analysing Q-PCR results e.g. by using two or more analytical approaches to validate conclusions. In our view a common effort should be made to standardise methods for analysis and validation of Q-PCR results.
journal_name
BMC Mol Bioljournal_title
BMC molecular biologyauthors
Skern R,Frost P,Nilsen Fdoi
10.1186/1471-2199-6-10keywords:
subject
Has Abstractpub_date
2005-04-26 00:00:00pages
10issn
1471-2199pii
1471-2199-6-10journal_volume
6pub_type
杂志文章abstract：BACKGROUND:Real-time RT-PCR is the recommended method for quantitative gene expression analysis. A compulsory step is the selection of good reference genes for normalization. A few genes often referred to as HouseKeeping Genes (HSK), such as ACT1, RDN18 or PDA1 are among the most commonly used, as their expression is a...
journal_title：BMC molecular biology
pub_type： 杂志文章
doi：10.1186/1471-2199-10-99
更新日期：2009-10-30 00:00:00
abstract：BACKGROUND:The enzyme glucose-6-phosphatase catalyzes the dephosphorylation of glucose-6-phosphatase to glucose, the final step in the gluconeogenic and glycogenolytic pathways. Expression of the glucose-6-phosphatase gene is induced by glucocorticoids and elevated levels of intracellular cAMP. The effect of cAMP in re...
journal_title：BMC molecular biology
pub_type： 杂志文章
doi：10.1186/1471-2199-6-2
更新日期：2005-01-19 00:00:00
abstract：BACKGROUND:Members of the Sox gene family isolated from both vertebrates and invertebrates have been proved to participate in a wide variety of developmental processes, including sex determination and differentiation. Among these members, Sox30 had been considered to exist only in mammals since its discovery, and its e...
journal_title：BMC molecular biology
pub_type： 杂志文章
doi：10.1186/1471-2199-11-98
更新日期：2010-12-11 00:00:00
abstract：BACKGROUND:We reported previously that the cardiac troponin I (cTnI) promoter drives cardiac-specific expression of reporter genes in cardiac muscle cells and in transgenic mice, and that disruption of GATA elements inactivates the cTnI promoter in cultured cardiomyocytes. We have now examined the role of cTnI promoter...
journal_title：BMC molecular biology
pub_type： 杂志文章
doi：10.1186/1471-2199-8-78
更新日期：2007-09-17 00:00:00
abstract：BACKGROUND:Transcription initiation by RNA polymerase II is unidirectional from most genes. In plants, divergent genes, defined as non-overlapping genes organized head-to-head, are highly represented in the Arabidopsis genome. Nevertheless, there is scarce evidence on functional analyses of these intergenic regions. Th...
journal_title：BMC molecular biology
pub_type： 杂志文章
doi：10.1186/1471-2199-10-95
更新日期：2009-10-16 00:00:00
abstract：BACKGROUND:Improving the treatment of renal cell carcinoma (RCC) will depend on the development of better biomarkers for predicting disease progression and aiding the design of appropriate therapies. One such marker may be fatty acid binding protein 7 (FABP7), also known as B-FABP and BLBP, which is expressed normally ...
journal_title：BMC molecular biology
pub_type： 杂志文章
doi：10.1186/1471-2199-12-31
更新日期：2011-07-19 00:00:00
abstract：BACKGROUND:Traumatic brain injury models are widely studied, especially through gene expression, either to further understand implied biological mechanisms or to assess the efficiency of potential therapies. A large number of biological pathways are affected in brain trauma models, whose elucidation might greatly benef...
journal_title：BMC molecular biology
pub_type： 杂志文章
doi：10.1186/1471-2199-9-62
更新日期：2008-07-08 00:00:00
abstract：BACKGROUND:cDNA-RDA is one of the subtractive cloning techniques used to isolate differentially expressed genes between two complex cDNA populations. In the present study we present a modification of the protocol described by Hubank and Schatz. RESULTS:In the post-hybridization mix, the 5'-ends of homoduplexes of inte...
journal_title：BMC molecular biology
pub_type： 杂志文章
doi：10.1186/1471-2199-3-6
更新日期：2002-04-24 00:00:00
abstract：BACKGROUND:GFG/NUDT is a nudix hydrolase originally identified as the product of the fibroblast growth factor-2 antisense (FGF-AS) gene. While the FGF-AS RNA has been implicated as an antisense regulator of FGF-2 expression, the expression and function of the encoded GFG protein is largely unknown. Alternative splicing...
journal_title：BMC molecular biology
pub_type： 杂志文章
doi：10.1186/1471-2199-9-10
更新日期：2008-01-23 00:00:00
abstract：BACKGROUND:Modulation of chromatin structure has emerged as a critical molecular device to control gene expression. Histones undergo different post-translational modifications that increase chromatin accessibility to a number of regulatory factors. Among them, histone ubiquitination appears relevant in nuclear processe...
journal_title：BMC molecular biology
pub_type： 杂志文章
doi：10.1186/1471-2199-10-55
更新日期：2009-06-05 00:00:00
abstract：BACKGROUND:The mRNA translation initiation region (TIR) comprises the initiator codon, Shine-Dalgarno (SD) sequence and translational enhancers. Probably the most abundant class of enhancers contains A/U-rich sequences. We have tested the influence of SD sequence length and the presence of enhancers on the efficiency o...
journal_title：BMC molecular biology
pub_type： 杂志文章
doi：10.1186/1471-2199-8-100
更新日期：2007-10-31 00:00:00
abstract：BACKGROUND:Wnt signaling is mediated through 1) the beta-catenin dependent canonical pathway and, 2) the beta-catenin independent pathways. Multiple receptors, including Fzds, Lrps, Ror2 and Ryk, are involved in Wnt signaling. Ror2 is a single-span transmembrane receptor-tyrosine kinase (RTK). The functions of Ror2 in ...
journal_title：BMC molecular biology
pub_type： 杂志文章
doi：10.1186/1471-2199-9-11
更新日期：2008-01-23 00:00:00
abstract：BACKGROUND:MicroRNAs (miRNAs) are short non-protein-coding RNA species that have a regulatory function in modulating protein translation and degradation of specific mRNAs. MicroRNAs are estimated to target approximately 60% of all human mRNAs and are associated with the regulation of all physiological processes. Simila...
journal_title：BMC molecular biology
pub_type： 杂志文章
doi：10.1186/s12867-016-0057-9
更新日期：2016-02-09 00:00:00
abstract：BACKGROUND:The human Sex Hormone Binding Globulin (SHBG) gene, located at 17p13.1, comprises, at least, two different transcription units regulated by two different promoters. The first transcription unit begins with the exon 1 sequence and is responsible for the production of plasma SHBG by the hepatocytes, while the ...
journal_title：BMC molecular biology
pub_type： 杂志文章
doi：10.1186/1471-2199-10-59
更新日期：2009-06-17 00:00:00
abstract：BACKGROUND:In spite of large intergenic spaces in plant and animal genomes, 7% to 30% of genes in the genomes encode overlapping cis-natural antisense transcripts (cis-NATs). The widespread occurrence of cis-NATs suggests an evolutionary advantage for this type of genomic arrangement. Experimental evidence for the regu...
journal_title：BMC molecular biology
pub_type： 杂志文章
doi：10.1186/1471-2199-9-6
更新日期：2008-01-14 00:00:00
abstract：BACKGROUND:Initiation of eukaryotic DNA replication involves many protein-protein and protein-DNA interactions. We have previously shown that 14-3-3 proteins bind cruciform DNA and associate with mammalian and yeast replication origins in a cell cycle dependent manner. RESULTS:By expressing the human 14-3-3epsilon, as...
journal_title：BMC molecular biology
pub_type： 杂志文章
doi：10.1186/1471-2199-8-27
更新日期：2007-04-12 00:00:00
abstract：BACKGROUND:Although much is known about molecular mechanisms that prevent re-initiation of DNA replication on newly replicated DNA during a single cell cycle, knowledge is sparse regarding the regions that are most susceptible to re-replication when those mechanisms are bypassed and regarding the extents to which check...
journal_title：BMC molecular biology
pub_type： 杂志文章
doi：10.1186/1471-2199-8-119
更新日期：2007-12-21 00:00:00
abstract：BACKGROUND:Physical exercise can improve brain function by altering brain gene expression. The expression mechanisms underlying the brain's response to exercise still remain unknown. miRNAs as vital regulators of gene expression may be involved in regulation of brain genes in response to exercise. However, as yet, very...
journal_title：BMC molecular biology
pub_type： 杂志文章
doi：10.1186/s12867-019-0120-4
更新日期：2019-01-15 00:00:00
abstract：BACKGROUND:Single-stranded DNA-binding proteins are essential cellular components required for the protection, metabolism and processing of single-stranded DNA. Human single-stranded DNA-binding protein 1 (hSSB1) is one such protein, with described roles in genome stability maintenance and transcriptional regulation. A...
journal_title：BMC molecular biology
pub_type： 杂志文章
doi：10.1186/s12867-016-0077-5
更新日期：2016-12-09 00:00:00
abstract：BACKGROUND:Many studies of the eukaryotic transcription mechanism and its regulation rely on in vitro assays. Conventional RNA polymerase II transcription assays are based on radioactive labelling of the newly synthesized RNA. Due to the inefficient in vitro transcription, the detection of the RNA involving purificatio...
journal_title：BMC molecular biology
pub_type： 杂志文章
doi：10.1186/1471-2199-15-7
更新日期：2014-04-03 00:00:00
abstract：BACKGROUND:Transient Receptor Potential Canonical 1 (TRPC1) is a widely-expressed mammalian cationic channel with functional effects that include stimulation of cardiovascular remodelling. The initial aim of this study was to investigate variation in TRPC1-encoding gene transcripts. RESULTS:Extensive TRPC1 transcript ...
journal_title：BMC molecular biology
pub_type： 杂志文章
doi：10.1186/1471-2199-12-30
更新日期：2011-07-12 00:00:00
abstract：BACKGROUND:Senescence marker protein-30 (SMP30), whose expression declines during aging in rat liver, has been proposed as an important aging marker. Besides apoptosis, SMP30 also protects cells against various other injuries by enhancement of membrane calcium-pump activity. The mechanism of this differential gene expr...
journal_title：BMC molecular biology
pub_type： 杂志文章
doi：10.1186/1471-2199-9-43
更新日期：2008-04-29 00:00:00
abstract：BACKGROUND:Heart and lung transplantation is frequently the only therapeutic option for patients with end stage cardio respiratory disease. Organ donation post brain stem death (BSD) is a pre-requisite, yet BSD itself causes such severe damage that many organs offered for donation are unusable, with lung being the orga...
journal_title：BMC molecular biology
pub_type： 杂志文章
doi：10.1186/1471-2199-10-72
更新日期：2009-07-23 00:00:00
abstract：BACKGROUND:Pseudo-nitzschia multiseries Hasle (Hasle) (Ps-n) is distinctive among the ecologically important marine diatoms because it produces the neurotoxin domoic acid. Although the biology of Ps-n has been investigated intensely, the characterization of the genes and biochemical pathways leading to domoic acid bios...
journal_title：BMC molecular biology
pub_type： 杂志文章
doi：10.1186/1471-2199-14-25
更新日期：2013-11-01 00:00:00
abstract：BACKGROUND:Among the three functions of DNA, mammalian replication and transcription can be subject to epigenetic imprinting specified by the parental origin of chromosomes, and although there is suggestive indication that this is also true for meiotic recombination, no definitive evidence has yet been reported. RESUL...
journal_title：BMC molecular biology
pub_type： 杂志文章
doi：10.1186/1471-2199-10-43
更新日期：2009-05-13 00:00:00
abstract：BACKGROUND:SRY is the pivotal gene initiating male sex determination in most mammals, but how its expression is regulated is still not understood. In this study we derived novel SRY 5' flanking genomic sequence data from bovine and caprine genomic BAC clones. RESULTS:We identified four intervals of high homology upstr...
journal_title：BMC molecular biology
pub_type： 杂志文章
doi：10.1186/1471-2199-9-85
更新日期：2008-10-14 00:00:00
abstract：BACKGROUND:Core canonical histones are required in the S phase of the cell cycle to pack newly synthetized DNA, therefore the expression of their genes is highly activated during DNA replication. In mammalian cells, this increment is achieved by both enhanced transcription and 3' end processing. In this paper, we descr...
journal_title：BMC molecular biology
pub_type： 杂志文章
doi：10.1186/s12867-018-0110-y
更新日期：2018-07-27 00:00:00
abstract：BACKGROUND:Krüppel-like factors (KLFs) are a group of master regulators of gene expression conserved from flies to human. However, scant information is available on either the mechanisms or functional impact of the coupling of KLF proteins to chromatin remodeling machines, a deterministic step in transcriptional regula...
journal_title：BMC molecular biology
pub_type： 杂志文章
doi：10.1186/1471-2199-15-10
更新日期：2014-05-25 00:00:00
abstract：BACKGROUND:Visceral leishmaniasis caused by members of the Leishmania donovani complex is often fatal in the absence of treatment. Research has been hampered by the lack of good laboratory models and tools for genetic manipulation. In this study, we have characterised a L. infantum line (JPCM5) that was isolated from a...
journal_title：BMC molecular biology
pub_type： 杂志文章
doi：10.1186/1471-2199-7-42
更新日期：2006-11-13 00:00:00
abstract：BACKGROUND:Nucleosomes are nucleoproteic complexes, formed of eight histone molecules and DNA, and they are responsible for the compaction of the eukaryotic genome. Their presence on DNA influences many cellular processes, such as transcription, DNA replication, and DNA repair. The evolutionarily conserved histone vari...
journal_title：BMC molecular biology
pub_type： 杂志文章
doi：10.1186/1471-2199-10-18
更新日期：2009-03-04 00:00:00
