B2m housekeeping gene At stage 3, B2M (Beta-2-Microglobulin) is a Protein Coding gene. The levels of six housekeeping genes (including 18 SrRNA, GAPDH, ACTB, RPL13A, B2M and TBP) in the skin tissue of fine-wool sheep were estimated by SYBR Green I which belongs to a method of real-time fluorescence quantitative RT-PCR expression. Expression stability (M) was assessed using normalization Background Research using the zebrafish model has experienced a rapid growth in recent years. 91 respectively) as compared to hypoxia (Average SD: 3. (GUSB); 0. Variations in the gene expression of common endogenous reference housekeeping genes (HKGs) used in data We studied five housekeeping genes preselected to the 96-well kit of human RT2 Profiler PCR Arrays (Qiagen, CA, USA) (ACTB, B2M, RPLP0, HPRT1, GAPDH) and two genes selected based on literature For accurate and reliable gene expression analysis, normalization of gene expression data against housekeeping genes (reference or internal control genes) is required. Pkd1, polycystic kidney disease 1 gene. Workflow diagram illustrating the strategy used to identify the best housekeeping normalizer for RT-qPCR studies in Pkd1-deficient kidneys. Best 1471-2172-11- MHC I and b2m genes are both expressed in in human as well as mice during the developmental stages of cerebellar cortex; An association between serum B2M and non-Hodgkin lymphoma prognosis. S3 Figure: melt curves of the ten candidate housekeeping genes. B2M protein expression summary. So far, reports The present study aimed to examine 10 housekeeping genes (HKGs), including 18s ribosomal RNA (18S), glyceraldehyde‑3‑phosphate dehydrogenase (GAPDH), ribosomal protein large P0 (RPLP0), β‑actin (ACTB), peptidylprolyl isomerase A (PPIA), phosphoglycerate kinase‑1 (PGK1), β‑2‑microglobulin (B2M), ribosomal protein LI3a (RPL13A), hypoxanthine phosphoribosyl . We selected group 2 genes as they The ΔCt values were determined by: Ct(target gene)−Ct(housekeeping gene) or Ct(target gene)−mean[Ct(housekeeping gene 1);Ct(housekeeping gene 2)]. Ten housekeeping genes were selected from commonly used reference genes (ABL1, ACTB, B2M, GAPD, GNB2L1, HRPT1, PBGD, RPL32, TBP, and TUBB). We sought to identify candidate control genes by analyzing seven functionally distinct housekeeping genes (B2M, GAPDH, HMBS, HPRT, SDHA, TBP, YWHAZ) for their expression stability and level in the placenta. Although the order of stability of the studied genes differed slightly among the 3 applets, the top 2 ranked genes exhibited Eight potential housekeeping genes were evaluated for expression level and stability among the 16-paired samples. View. Box-and-whisker plot showing Cq values of 12 housekeeping genes examined in the Developmental panel, displayed in different age groups (a) or displayed as all samples in the panel (c). Five potential housekeeping genes, GAPDH, RPNI, PUM1, B2M, and PMM1, were assessed for their expression stability on qPCR using Sybr Green Chemistry and melt curve analysis. 79 and 2. B2M B2m ++ to +++ ++ to +++ Tyrosine 3 In GeNorm all tested genes reached the threshold, and the algorithm revealed that two housekeeping genes are sufficient for a subsequent gene expression analysis (Fig. For accurate and reliable gene expression analysis, normalization of gene expression data against housekeeping genes (reference or internal control genes) is required. Our results showed that B2m, Gapdh, and Hprt, or a combination of B2m/Gapdh and B2m/Hprt, were the most stably expressed genes or gene pairs. The ranking by RefFinder that provided a robust The housekeeping gene expression levels are fine-tuned to meet the metabolic requirements in various tissues. Likewise Ideal housekeeping genes must have stable tissue expression, but few have been specifically studied in the placenta. ACTB, GAPDH, HPRT1, and B2M) vary considerably under different experimental Housekeeping genes considered suitable for qRT-PCR normalization are the ones present in all nucleated cell types, necessary for basic cell survival and considered stable in various tissues. Cq values of 12 housekeeping genes examined in the Light/Dark rearing panel, displayed in different The lower deviation was observed when HMBS and B2M were used as housekeeping genes for ACTB and 18S rRNA expression analysis in 3T3-L1 cells under normoxia (Average SD: 2. At stage 2, the most stable gene was EIF3K and B2M was the least stable (Fig. In parallel, the study This study provides data that can be expected to aid zebrafish researchers in their initial choice of housekeeping genes for future studies, but underlines the importance of further validating housekeeping genes for each new experimental paradigm and fish species. Hypertensive patients with hyperuricemia show higher plasma concentrations of beta-2-microglobulin (B2M), compared to patients without uric acid metabolic Candidate genes for expression studies. It is very stable in T cells in different activation states, and has low variance between subjects. Therefore, choosing housekeeping genes is an essential step for the standardizing of RT-qPCR since using unsuitable reference genes can lead to discrepant and unreliable data (Cai et al. May 2013; Housekeeping genes. A – highlights a subset of candidate housekeeping genes including B2M, GAPDH and SDHA. 8A and B, for both chondrogenesis and osteogenesis the use of the most unstable B2M gene gave rise to a dramatic effect on the correct housekeeping genes (Actb, Actg1, B2m, Gapdh, Hprt, Pgam1 and Ppia) in kidney tissues from mouse models orthologous to ADPKD, including a cystic mice (CY) 10–12 weeks old (Pkd1 ox/ox : Nestin cre the expression levels of ALP normalized using the most stable reference genes (B2M/ HMBS/HPRT1) differed greatly from the expression levels obtained when the data were nor-malized using the least stable genes (GAPDH, B-actin, and 18S). Gene symbols and their full names, gene accession numbers as well as functions are listed in Table 1. This gene encodes a serum protein found in association with the major histocompatibility complex (MHC) class I heavy cha in on the surface of nearly all nucleated cells. The selection of five housekeeping genes (HPRT, GAPDH, 18S, UBC, B2M) is based on the literature (Table 2). Methods and results: Expression of six housekeeping genes (HKG's): Glyceraldehyde-3-phosphodehydrogenase (GAPDH), β-actin (ACTB), beta-2-microglobulin (B2M), 18S ribosomal RNA (18S rRNA), peptidyl-propyl-isomerase A (PPIA), and ribosomal protein L13 (RPL-13) during two dimensional (2D) culture, and alginate-carboxymethylcellulose scaffold Scientific Reports - Screening and validating the optimal panel of housekeeping genes for 4T1 breast carcinoma and metastasis studies in mice. In all algorithms, Tubb2a was ranked as the least stable candidate in the liver. EE1A and B2M were abundantly expressed genes in all considered cells presenting average Ct values below 14 and 15 cycles, respectively Background Assessing an optimal reference gene as an internal control for target gene normalization is important during quantitative real time polymerase chain reaction (RT-qPCR) of three dimensional (3D) cell culture. Especially, gene profiling of cancer cells under a complex 3D microenvironment in a polymer scaffold provides a deeper understanding of tumor The expression stability of 14 housekeeping genes (HKGs; ACTB, B2M, G6PD, GAPDH, GUSB, HMBS, HPRT1, IPO8, PGK1, PPIA, TBP, TFRC, UBC, and YWHAZ) after irradiation was evaluated using RefFinder Since the current study also observed fluctuations in expression in frequently used housekeeping genes, including 18S rRNA, B2M, CYPA, GAPDH, HMBS, HPRT1, SDHA and TBP, it might be rather recommended to use ACTB, GUSB and RPL13A as housekeeping genes for lung tissues in studies of similar design but also to analyze the stability of housekeeping Results indicate that PPIB (cyclophilin B), B2M (beta-2-microglobulin) and PPIA (cyclophilin A) as the best combination for gene expression normalization in this context. (II) Duplicative regions, may bias expression-level measurement. According to BestKeeper, B2M/ But use of unconfirmed housekeeping genes (HKGs) could lead to misinterpretation of the expression of genes of interest (GOI). A list of 7780 genes was first obtained by selecting genes with low 0’s proportion and coefficient of variation (CV). 7 At the molecular biology level, the expression and stability of ve commonly used housekeeping genes B2M, RPL13A, PPIA, HMBS, and GAPDH, were considered for comparing and nding out ideal reference gene. Ct values <40 were only used for calculation of the PCR efficiency. To evaluate potential expression differences of Actb, Actg1, B2m, Gapdh, Hprt, Pgam1, Ppia and Stat3 among the sample groups, the Kruskal–Wallis with Dunn’s post-hoc test, followed by FDR Background Real-time quantitative polymerase chain reaction (RT-qPCR) is a widely-used standard assay for assessing gene expression. The protein has a predominantly beta-pleated sheet structure that can form amyloid fibrils in some pathological conditions. For relative quantification of gene expression, it is crucial to select an appropriate housekeeping gene as a reference. There is no evidence for the existence of B2M pseudogenes. The corresponding Figure 1: Problems occuring in Housekeeping Gene detection. The expression of the 4 indicated housekeeping genes was examined using single-cell RT-PCR in fresh HSCs (CD150 + CD34 − KSL cells). (A) Frequency of HSCs in which the expression of The aim of this study was to better understand these structures by focusing on the choice of the best housekeeping gene (HKG) to perform accurate molecular analysis on such a heterogeneous system. Although real-time reverse transcription PCR (QPCR), normalized to an internal reference ("housekeeping") gene, is a frequently used method for quantifying gene expression changes in zebrafish, many commonly used housekeeping genes are known to vary with The expression stability of 14 housekeeping genes (HKGs; ACTB, B2M, G6PD, GAPDH, GUSB, HMBS, HPRT1, IPO8, PGK1, PPIA, TBP, TFRC, UBC, and YWHAZ) after irradiation was evaluated using RefFinder Normalize real time PCR results to multiple housekeepers rather than one for more accurate quantitation Perform up to 1000 assays (based on 20 µl assay volume) for each housekeeping gene Includes primer sets for ACTB, B2M, GAPD, GUSB, HPRT1, PGK, PP1A, RPL13A, TBP and Data normalisation in real-time RT-PCR is a further major step in gene quantification analysis (Bustin 2002, Pfaffl 2001). B – shows comparisons of the Constitutive or “housekeeping” (HK) genes are those whose products are involved in basic cell maintenance biological functions. 1 (ABL). Five potential housekeeping genes, GAPDH, RPNI, PUM1, B2M, and PMM 1, were assessed for their. Human PBGD We evaluated the expressional changes of eight well-known housekeeping genes (i. Among tested housekeeping genes, PPIA and RPS13, especially in combination, proved best suitable to normalize gene expression in ccRCC tissues as compared to classical reference genes such as beta-Actin, GAPDH, 18S or B2M. 1 (G6PDH), 26. Trauma triggers critical molecular and cellular signaling cascades that drive biological outcomes and recovery. The identification of the best housekeeping genes (HKG) is a critical step for studying the mRNA expression of a target gene. It is known that commonly used housekeeping Several genes have been used as housekeeping genes, including β2-microglobulin (B2M), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), β-actin (ACTB), hypoxanthine phosphoribosyl transferase (HPRT) and ribosomal RNA (18 s and 28 s rRNA) [18,21]. mRNA isolated Mrpl32 was the best HKGs in the liver and kidney and Tubb2a|Trfc and B2m must be avoided, also called housekeeping or internal control genes. 5F). (I) Genes having several splice variants could have different expression levels for different parts of the gene. Housekeeping genes are genes that are essential for the maintenance of cellular function, such as cell metabolism and structural gene . ΔCt variability in candidate housekeeping gene comparisons are shown as medians (lines), 25 th percentile to the 75 th percentile (boxes) and ranges (whiskers) for 31 human reticulocyte samples. 16 * Ratio of arbitrary fl uorescent A common method is to report the interest gene expression relative to an internal control, usually a housekeeping gene (HKG), which should be constant in cells independently of experimental conditions. HKGs suitable for quantitative B2m NM_009735 29 hamster probes 38. It is known that commonly used housekeeping genes (e. The 10 genes investigated in this assay are shown in Table 1. Based on this study we recommend the use of at least two of the following genes: RPII, B2M, RPL13A. S1 Table: expression stability values of ten candidate housekeeping genes in L02 analyzed by NormFinder. Comparable results were also seen in the However, beta-Actin, and TBP were identified as the most stable housekeeping genes (HKGs) to carry out genetic based expression analysis of all samples associated with Covid-19. In contrast The objective of this study was to determine hypoxia-stable housekeeping genes (HKG) for quantitative real-time PCR (qRT-PCR) in human chondrocytes cultured in 21%, 5% and 1% oxygen by geNorm and NormFinder analyses. e. RT-qPCR data requires reference genes for normalization to make the results comparable. 5 in the normal samples compared to 25. Housekeeping genes are essential for maintaining basic cellular functions and are generally expressed at constant levels across different tissues and conditions. 18 respectively). Diseases associated with B2M include Immunodeficiency 43 and Amyloidosis, Hereditary Systemic 6. The list B2M was previously investigated as a candidate reference gene when examined in reprogrammed iPSC or EC and was ranked between four to eight of a panel of 10 housekeeping genes 18,29, 38. the stability and suitability of 11 frequently used housekeeping genes, namely 18S rRNA, ACTB, B2M, CYPA, GADPH, GUSB, HMBS, HPRT1, RPL13A, SDHA and TBP in 36 lung tissues isolated from either wild-type (WT) mice or A housekeeping gene is defined as a gene that encodes proteins essential for maintaining basic cell functions. The present study, therefore, aims to fill the void by investigating the gene expression of twelve reference genes that were previously identified as stable genes by various studies [25][26][27 The expression stability was assessed using five computational algorithms: BestKeeper, geNorm, NormFinder, DeltaCt, and RefFinder. b2m, and elfa as most stable during development and across tissue types Instability of housekeeping genes (HKG), supposedly unregulated and hence used as normalizers, may dramatically change conclusions of quantitative PCR experiments. An important paralog of this gene is HLA The choice of housekeeping genes for the study. The average Ct values significantly decreased over time in B2M gene and 18S showed a little bit higher Ct value on PND1, S2 Figure: agarose gel (1%) electrophoresis of the ten candidate housekeeping genes. HKGs are often used to correct sample-to-sample variations, including the amount and integrity of materials, enzymatic efficiency, cDNA sample loading, and differences between Background RT-PCR has been widely used for the analysis of gene expression in many systems, including tumor samples. Validation of Stable Housekeeping Genes for Quantitative Real Time PCR in Golden Syrian Hamster Eef2, Ywhaz, Rps18, Hprt1, Tubb, Rpl13a, Nono, and B2m, in hamster pancreas at different time B2M, HPRT1, and TBP genes are recommended for NCS treatment, while 18S rRNA, B2M, and PPIA genes can be used as suitable internal controls in RT-qPCR experiments for ActD treatment. The reliability of any relative RT-PCR experiment can be improved by including an invariant endogenous In the present study, we have identifi ed species-specifi c housekeeping genes (HKGs) for Chinese Hamster Ovary (CHO) cells using data from microarray gene expression profi ling. 1-10 represent ACTB, B2M, GAPDH, TUBB2a, HPRT1, SDHA, TBP, YWHAZ, CYC1, and GUSB, respectively. We concluded that B2M/ housekeeping genes should be well assessed to circumvent problems for data Reliability of selected housekeeping genes. HKG should not be co-regulated with the target gene or influenced by the experimental conditions (Lemma et al. B2M, HPRT1, ACTB, 18 S, and GAPDH genes The best combination of HKGs for the RKO-AS45-1 was B2M/GAPDH. To proceed with appropriate comparisons, we included housekeeping genes selected from the available literature (Actb, Actg1, B2m, Gapdh, Hprt, Pgam1 and Ppia). Among its related pathways are SARS-CoV-2 Infection and Infectious disease. In the context of Q-PCR, it is used as a reference to normalize target gene expression and account for variations in RNA content during the experimental process. , 2019 Descriptive statistics of 12 candidate reference genes in two sets of choroid plexus samples. , 18S rRNA, B2M, eEF1α1, GAPDH, GUSB, HPRT1, PPIA, and TBP) following treatment with the DNA-damaging agents that FAM-MGB coupled Taqman probes and housekeeping genes used for quantitative RT-PCR. 8 (PBGD) and 27. g. 65 and 1. Gene Ontology (GO) annotations related to this gene include identical protein binding. They belong to different functional classes and are reported to be used in genomic studies of immune cells. ACTB was also used as a reference gene to confirm results Which housekeeping gene is the best choice for RT-qPCR analysis in mice fed with a high-fat diet? Studies in the liver, kidney, pancreas, and intestines GeNorm, and Delta-Ct, while BestKeeper showed B2m and Ppia as more stable genes. In contrast, Actb and Rpl13a were the least stably expressed. 23% 62. Housekeeping genes (HKG) serve as a common denominator to which target gene expression is normalized, making the identification of stable HKG for real time RT-PCR a critical requirement for accurate and biologically meaningful analysis of gene expression. All of these genes were analyzed for their stability in 20 rectal tumors and six normal rectal samples through qPCR as part of the screening phase through Bestkeeper software. , 2015) and about The housekeeping gene B2M was used for normalization since this reference gene has been shown to have stable expression in PBMCs [34, 35]. Gene Symbol Gene Name RefSeq Accession # B2M: Beta-2-microglobulin: NM_004048. Biochemical studies on transcription initiation of the housekeeping gene promoters have been difficult, partly due to the less-characterized promoter motifs and transcription initiation process. We show that HMBS and B2M are stably expressed under hypoxia associated obesity like conditions and could be considered as internal controls for related studies in future. In this study, we investigated 70 genes to provide a comprehensive overview of the most of housekeeping genes used in literature (Table 1). All of these genes Flowchart of a novel identification strategy for housekeeping genes (nHKGs) in breast cancer. The Mann–Whitney test was performed for each reference gene. Similar results were obtained when the study was conducted using RAW264. As shown in Fig. , 2016). This result is coherent since B2M as part of MHC I Here, we determined that beta2-microglobulin (B2m) is an appropriate reference gene for analyzing mouse HSCs by a novel method using single-cell RT-PCR. The relative transcriptional level of the two ductal markers EpCAM and SOX9 in each sample condition considered was normalized with the best and the Housekeeping Gene for Real-Time Gene Proling of Hepatocellular Carcinoma Cells Cultured in Three- 18S rRNA and B2M to be stable among selected referral HKG candidates and considered them as Selection of Housekeeping Genes. However, the appropriateness of these genes should be validated for each Due to different algorithms in three tools, there was slight discrepancy in the stability ranking of candidate genes. 97% 544. We first studied the gene expression levels of 10 housekeeping genes (designated group 1) using a commercially available assay (TaqMan ® human endogenous control plate; Applied Biosystems, Foster City, CA, USA) (Citation 13). The mean CP of B2M was 13. My new favorite housekeeping gene is OAZ1, which I common housekeeping genes, B2M, GAPDH, GUSB, HMBS, and HPRT1, to compare their stability with the 15 candidate genes. B2M exceeded the level of the other genes by far. Data were analyzed using four practical algorithmic approaches, including Norm Finder, GeNorm, Best Keeper, and the Delta Ct for reference gene The best gene identified by this program was B2M , and the best combination of two genes was B2M and RPLP0. 5, 4, 8, and 12hpi in LTC with a SPPV-WS Jaipur was examined under this study using four different frequently used programs such as the Normfinder, BestKeeper, geNorm, and ΔCT in RefFinder. 2 Keywords: qPCR, Reference gene, Housekeeping gene, Gene expression, Rat, Toxicology, qRT-PCR, Testis, Liver, Prostate Introduction Despite the advent of high-throughput methods such as RNA-sequencing to measure transcript abundance in cells and tissues, quantitative RT-PCR (RT-qPCR) remains the method of choice for many, particularly when only We sought to identify candidate control genes by analyzing seven functionally distinct housekeeping genes (B2M, GAPDH, HMBS, HPRT, SDHA, TBP, YWHAZ) for their expression stability and level in the placenta. However, GAPDH has been shown to be up-regulated in Housekeeping genes (HKGs) are essential for basic maintenance of a variety of cellular processes. 128 for B2M respectively. expression stability on qPCR using Sy br Green Chemistry and melt curve an alysis. According to GeNorm, B2M/HMBS/HPRT1 were the optimal housekeeping genes in the duo- denum, jejunum, ileum, and colon as well as the whole GIT. 3 B). The results indicated that B2M (P0 vs P5), ACTB (P0 vs P5 and P2 vs P5), GAPDH (P0 B2M is now among our favorite candidates. These genes were chosen because they have different Housekeeping genes. 02 Gapdh NM_008084 61 hamster probes 44. Expression stability of 15 HKG (ACTB, B2M, GAPDH, GUSB, HPRT1, IPO8, MRPL19, PG At present, transcription analysis of gene expression commonly uses a single housekeeping gene as control for normalization. For example, B2M was the best suitable reference gene in NCM460, HT29, HCT116 Raw RT-qPCR data in renal tissue were obtained for the Actb, Actg1, B2m, Gapdh, Hprt, Pgam1 and Ppia candidate housekeeping genes for each of the Pkd1-deficient mouse models. B2M: Human B2M : ABIN3340183: Murine B2M : ABIN3275845: Rattus B2M : ABIN3352421: Tyrosine 3 The present data indicate that TBP and B2M are appropriate housekeeping genes for normalization of transcript abundance measured by real-time RT-PCR in granulosa cells subjected to different plating densities, oxygen concentrations and FSH stimulation. Raw RT-qPCR data in renal At present, use of the so-called housekeeping genes (HKGs), endogenous control genes or reference genes, is the gold standard to normalize mRNA fractions of interest. . e specicity and the amplication range of the primers that were designed for These genes are thus distributed into different expression level categories, the genes with high abundance, RN18S1 and GAPDH; genes with moderate abundance, HPRT and B2M; and genes with low abundance, TBP and RPL13A. 0’s proportion is defined as the proportion of different cell lines in which a given gene is not expressed; and CV is the ratio of the standard variation to the mean. A set of housekeeping genes such as B2M, HMBS, HPRT1, HSP-90 (HSPCA), POLR2A, 18s_RNA, and GAPDH at 0. We identified a panel of housekeeping genes that could be utilized in various conditions using iPSC and iPSC-derived endothelial cells as well as genetically modified iPSC Several genes have been used as housekeeping genes, including β2-microglobulin (B2M), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), β-actin (ACTB), hypoxanthine phosphoribosyl transferase (HPRT) and ribosomal Using this simple approach, GAPDH was found to be the most suitable housekeeping gene for expression studies in reticulocytes while the Here, we determined that beta2-microglobulin (B2m) is an appropriate reference gene for analyzing mouse HSCs by a novel method The objective of this study is to identify and validate the most appropriate housekeeping genes for RNA expression analysis of human primary peripheral blood mononuclear cells (PBMCs) in In this study, we validated the expression of 12 well established housekeeping genes in ChP in 2 independent experimental paradigms by using popular stability testing In this work, we evaluated nineteen genes of different functional classes using Real Time Human Reference Gene Panel (Roche Applied Sciences), to identify the internal Thus, B2M is the only one to complete all the recommendations and is the best candidate for internal control of gene expression in RT-qPCR. mRNA isolated from 20 placentae was analyzed for gene expression using RT-PCR. In summary, this is the first systematic study using a U2OS cell culture system that offers convincing evidence for housekeeping gene selection following treatment Selection of Interest Housekeeping Genes . The effect of bowel inflammation on HKG remains unknown. 965 for tyrosine 3-monooxygenase (YWHAZ) and 1. B2m expression could be stably detected in uncultured, fresh HSCs. Through this work, we studied the expression profile of four commonly used housekeeping genes namely ACTB, 18S rRNA, HMBS, and B2M in hypoxia associated obesity like conditions. GAPDH (Glyceraldehyde-3-phosphate dehydrogenase) has been frequently considered as a constitutive housekeeping gene and used to normalize changes in specific gene expression. It has long been recognized that while some genes show a variable degree of case-specific expression (developmental age, tissue, normal or pathological conditions), others appear to be transcribed at a relatively stable rate across In summary, GAPDH, ACTB, RPLP0, HPRT1, and B2M are commonly used housekeeping genes for qPCR experiments in HEK293FT cells. DAP12 inhibition of LPS signaling in macrophages is mediated by DOK3 (P4147) Article. Selection of stable housekeeping genes (HKGs) is very important for accurate calculation of relative expression levels of target genes by quantitative real-time polymerase chain reaction (qRT-PCR). B2M B2m ++ to +++ ++ to +++ Tyrosine 3 Housekeeping genes are commonly used as a reference [2, 3]. Therefore, the selected reference gene should be highly stable in its expression throughout the experimental datasets. Clonally sorted The filtered cells were highly homogenous, as judged by that fact that >90% of cells expressed the B2M gene (a typical housekeeping gene of hematopoietic cells) (Matsuzaki et al. ΔCt approach to housekeeping gene selection.
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