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信號通路基因表達qPCR array—RealTimePrimers

技術文章

基于SYBR Green的實時定量PCR依舊是當前多數研究人員所依賴的價格相對低廉、的基因表達測定方法,RealTimePrimers致力于為科研工作者提供價格低廉,經高度優化的實時定量PCR引物系列,結合高質量的模板和試劑,RealTimePrimers產品將為您呈現清晰、的PCR實驗結果?,F今,越來越多的研究者選擇了RealTimePrimers產品。
 實時定量PCR產物的產物定量有兩種方式:使用已知濃度質粒DNA或對照基因的標準曲線來定量,和相對簡單的相對定量法。兩者都要保證擴增效率一致,但通常情況下,純靶標產物的擴增效率要高于情況復雜的混合物樣品,在實驗中尤其要注意避免酚污染和(對RNA反轉后定量)DNA污染等問題。
相對定量方法通過管家基因或對照基因來歸一化靶標基因,是更常用的定量方式。在這一過程中選擇合適的對照基因十分重要,通常需要一系列管家基因,以便綜合或是找到穩定的基因來完成這一工作。
 RealTimePrimers產品介紹:
1 信號通路PCR arrays (Pathway PCR arrays)
將信號通路上的基因集合形成arrays,一塊微板上含88個靶標基因外加8個管家基因(housekeeping gene),大幅度降低了實驗費用,提高了實驗效率。
已有信號通路產品如下:

Adaptive and Innate Immune Response

Adherin-Tight Junction

Adipogenesis

Amino acid metabolism

Androgen Signaling

Angiogenesis

Antigen Processing and Presentation

Apoptosis

Atherosclerosis

Autophagy

BMP Signaling

Breast Cancer Signaling

cAMP Signaling

Cancer Pathways

Celladhesion

Chromatin Regulation

Cell Cycle

CREB Signaling

Cytokines

Cytokine & chemokine Receptors

Cytoskeleton

DNA Damage Signaling

DNA Repair

Drug DetoxicationI

Drug Detoxication II

Drug TransportersI

EGFR Signaling

Epithelial-Mesenchymal Transition

Estrogen Signaling

Fatty Liver Signaling

FGFR Signaling

Focal Adhesion Signaling

Glucose Metabolism

GPCR Signaling

Heat Shock Proteins

Hedgehog Signaling

Hippo Signaling

Homeobox (HOX) Genes

Hypoxia Signaling

Insulin Signaling

Integrin Signaling

JAK/STAT Signaling Pathway

KINOME

Ligand Gated Ion Channels

Lipoprotein Signaling

MAPKinase Signaling Pathway

Neurotransmitter Signaling

NFkB Signaling Pathway

NGF Signaling

Nitric Oxide Signaling

Notch Signaling Pathway

Nuclear Receptorsand Coregulators

Oncogenes

Osteogenesis

Oxidative Stress

p53 Signaling Pathway

PDGF Signaling

PI3K-AKT Signaling

PKA Signaling Pathway

RAS Signaling

Rho GTPase Signaling

Signal Transduction

SMAD Signaling

Statin Signaling

Stem Cell

TGF beta Signaling Pathway

TNF Signaling

Toll-Like Receptor(TLR) Signaling

Tumor Invasion & Metastasis

Ubiquitination Pathway

VEGF Signaling

Wnt Signaling

CUSTOM Primer Libraries

 

 

 

以人繼發和先天免疫反應引物系列(Human Adaptive & Innate Immune Response Primer Library)為例,簡單給予描述:
信號通路PCR arrays以96孔微板形式提供,每板含88個引物對和8個管家基因引物對,只需加入cDNA模板和SYBR green master mix即可實驗。這一人繼發和先天免疫反應引物系列所含基因如下:
ADORA2A Adenosine A2a receptor
C5 Complement component 5
C8A Complement component 8, alpha polypeptide
CAMP Cathelicidin antimicrobial peptide
CASP1 Caspase 1, apoptosis-related cysteine peptidase (interleukin 1, beta, convertase)
CASP4 Caspase 4, apoptosis-related cysteine peptidase
CCL2 Chemokine (C-C motif) ligand 2
CCR3 Chemokine (C-C motif) receptor 3
CD14 CD14 molecule
CD1D CD1d molecule
CD55 CD55 molecule, decay accelerating factor for complement (Cromer blood group)
CHUK Conserved helix-loop-helix ubiquitous kinase
COLEC12 Collectin sub-family member 12
CRP C-reactive protein, pentraxin-related
CXCR4 Chemokine (C-X-C motif) receptor 4
CYBB Cytochrome b-245, beta polypeptide
DEFB4 Defensin, beta 4
DMBT1 Deleted in malignant brain tumors 1
FN1 Fibronectin 1
HMOX1 Heme oxygenase (decycling) 1
IFNA1 Interferon, alpha 1
IFNB1 Interferon, beta 1, fibroblast
IFNG Interferon Gamma
IFNGR1 Interferon gamma receptor 1
IFNGR2 Interferon gamma receptor 2 (interferon gamma transducer 1)
IKBKB Inhibitor of kappa light polypeptide gene enhancer in B-cells, kinase beta
IL10 Interleukin 10
IL12RB2 Interleukin 12 receptor, beta 2
IL1A Interleukin 1, alpha
IL1B Interleukin 1, beta
IL1F10 Interleukin 1 family, member 10 (theta)
IL1R1 Interleukin 1 receptor, type I
IL1R2 Interleukin 1 receptor, type II
IL1RAP Interleukin 1 receptor accessory protein
IL1RAPL2 Interleukin 1 receptor accessory protein-like 2
IL1RL2 Interleukin 1 receptor-like 2
IL1RN Interleukin 1 receptor antagonist
IL2 Interleukin 2
IL36RN Interleukin 36 receptor antagonist
IL36A Interleukin 36, alpha
IL37 Interleukin 37
IL36B Interleukin 36, beta
IL36G Interleukin 36, gamma
IL4 Interleukin 4
IL6 Interleukin 6 (interferon, beta 2)
IL8 Interleukin 8
IRAK1 Interleukin-1 receptor-associated kinase 1
IRAK2 Interleukin-1 receptor-associated kinase 2
IRF1 Interferon regulatory factor 1
LALBA Lactalbumin, alpha-
LBP Lipopolysaccharide binding protein
LTF Lactotransferrin
LY96 Lymphocyte antigen 96
LYZ Lysozyme
MAPK14 Mitogen-activated protein kinase 14
MAPK8 Mitogen-activated protein kinase 8
MIF Macrophage migration inhibitory factor (glycosylation-inhibiting factor)
MYD88 Myeloid differentiation primary response gene (88)
NCF4 Neutrophil cytosolic factor 4, 40kDa
NFKB1 Nuclear factor of kappa light polypeptide gene enhancer in B-cells 1
NFKB2 Nuclear factor of kappa light polypeptide gene enhancer in B-cells 2 (p49/p100)
NFKBIA Nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, alpha
NLRC4 NLR family, CARD domain containing 4
NOS2 Nitric oxide synthase 2, inducible
PGLYRP1 Peptidoglycan recognition protein 1
PGLYRP2 Peptidoglycan recognition protein 2
PGLYRP3 Peptidoglycan recognition protein 3
PPBP Pro-platelet basic protein (chemokine (C-X-C motif) ligand 7)
PROC Protein C (inactivator of coagulation factors Va and VIIIa)
PTAFR Platelet-activating factor receptor S100A12 S100 calcium binding protein A12
SERPINA1 Serpin peptidase inhibitor, clade A (alpha-1 antiproteinase, antitrypsin), member 1
SERPINE1 Serpin peptidase inhibitor, clade E (nexin, plasminogen activator inhibitor type 1), member 1
SFTPD Surfactant protein D
TGFB1 Transforming growth factor, beta 1
TLR1 Toll-like receptor 1
TLR10 Toll-like receptor 10
TLR2 Toll-like receptor 2
TLR3 Toll-like receptor 3
TLR4 Toll-like receptor 4
TLR6 Toll-like receptor 6
TLR8 Toll-like receptor 8
TLR9 Toll-like receptor 9
TNF Tumor necrosis factor
TNFRSF1A Tumor necrosis factor receptor superfamily, member 1A
TOLLIP Toll interacting protein
TRAF6 TNF receptor-associated factor 6
TREM1 Triggering receptor expressed on myeloid cells 1
ACTB Actin, beta
B2M Beta-2-microglobulin
GAPDH Glyceraldehyde-3-phosphate dehydrogenase
GUSB Glucuronidase, beta
HPRT1 Hypoxanthine phosphoribosyltransferase 1
Pgk1 Phosphoglycerate kinase 1
PPIA Peptidylprolyl isomerase A
RPL13A Ribosomal protein L13a

2. 實時定量PCR引物
Realtimeprimers提供您所研究基因經證實的引物系列,并提供引物序列信息和詳細的反應條件,可滿足1000次PCR反應所需。
這類引物系列包括人、小鼠和大鼠三個物種。

Human PCR Primer Sets

Rat PCR Primer Sets

Mouse PCR Primer Sets

以人PCR primer sets為例,包括:

 A2BP1   Human NA

 A2LP   Human NA

 A2M   Human alpha-2-macroglobulin

 A4GALT   Human alpha 1,4-galactosyltransferase

 AAAS   Human achalasia, adrenocortical insufficiency, alacrimia (Allgrove, triple-A)

()

 EPHX1   Human Epoxide hydrolase 1, microsomal

 MAPK3   Human mitogen-activated protein kinase 3

 PGR   Human Progesterone receptor

 18S   Human 18S Ribosomal RNA gene

 GUSB   Human Glucurondiase, beta

3.提供管家基因引物序列信息,夠1000次反應使用。通過比較多個看家基因表達,更好定量靶標基因的水平。
這類引物系列包括人、小鼠和大鼠三個物種。

Human Housekeeping Gene Primer Set

Mouse Housekeeping Gene Primer Set

Rat Housekeeping Gene Primer Set

以人housekeeping基因系列為例,包括基因:ACTB, B2M, GAPD, GUSB, HPRT1, PGK, PP1A, RPL13A, TBP和TFRC。

4. 基因家族(gene family)引物序列
將基因家族的成員匯總,提供已證實的引物序列,用于研究不同成員的表達差異,并提供引物的序列信息。
這類基因家族包括:

Annexin genes

BIRC (IAP) genes

Caspase genes

Chemokine genes

TNF genes

以人TNF基因家族為例,成員包括:TNF, LTA, LTB, TNFSF4, TNFSF5, TNFSF6, TNFSF7, TNFSF8, TNFSF9, TNFSF10, TNFSF11, TNFSF13, TNFSF13B, TNFSF14, TNFSF15, TNFSF18等
 

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