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信號(hào)通路基因表達(dá)qPCR array—RealTimePrimers
點(diǎn)擊次數(shù)：3795 發(fā)布時(shí)間：2018-7-31
基于SYBR Green的實(shí)時(shí)定量PCR依舊是當(dāng)前多數(shù)研究人員所依賴的價(jià)格相對(duì)低廉、的基因表達(dá)測(cè)定方法，RealTimePrimers致力于為科研工作者提供價(jià)格低廉，經(jīng)高度優(yōu)化的實(shí)時(shí)定量PCR引物系列,結(jié)合高質(zhì)量的模板和試劑，RealTimePrimers產(chǎn)品將為您呈現(xiàn)清晰、的PCR實(shí)驗(yàn)結(jié)果?，F(xiàn)今，越來越多的研究者選擇了RealTimePrimers產(chǎn)品。
實(shí)時(shí)定量PCR產(chǎn)物的產(chǎn)物定量有兩種方式：使用已知濃度質(zhì)粒DNA或?qū)φ栈虻臉?biāo)準(zhǔn)曲線來定量，和相對(duì)簡(jiǎn)單的相對(duì)定量法。兩者都要保證擴(kuò)增效率一致，但通常情況下，純靶標(biāo)產(chǎn)物的擴(kuò)增效率要高于情況復(fù)雜的混合物樣品，在實(shí)驗(yàn)中尤其要注意避免酚污染和(對(duì)RNA反轉(zhuǎn)后定量)DNA污染等問題。
相對(duì)定量方法通過管家基因或?qū)φ栈騺須w一化靶標(biāo)基因，是更常用的定量方式。在這一過程中選擇合適的對(duì)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(wěn)定的基因來完成這一工作。
RealTimePrimers產(chǎn)品介紹：
1 信號(hào)通路PCR arrays (Pathway PCR arrays)
將信號(hào)通路上的基因集合形成arrays，一塊微板上含88個(gè)靶標(biāo)基因外加8個(gè)管家基因(housekeeping gene)，大幅度降低了實(shí)驗(yàn)費(fèi)用，提高了實(shí)驗(yàn)效率。
已有信號(hào)通路產(chǎn)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(fā)和先天免疫反應(yīng)引物系列(Human Adaptive & Innate Immune Response Primer Library)為例，簡(jiǎn)單給予描述：
信號(hào)通路PCR arrays以96孔微板形式提供，每板含88個(gè)引物對(duì)和8個(gè)管家基因引物對(duì)，只需加入cDNA模板和SYBR green master mix即可實(shí)驗(yàn)。這一人繼發(fā)和先天免疫反應(yīng)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(shí)時(shí)定量PCR引物
Realtimeprimers提供您所研究基因經(jīng)證實(shí)的引物系列，并提供引物序列信息和詳細(xì)的反應(yīng)條件，可滿足1000次PCR反應(yīng)所需。
這類引物系列包括人、小鼠和大鼠三個(gè)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) |
等(略) |
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 |
等 |
3.提供管家基因引物序列信息，夠1000次反應(yīng)使用。通過比較多個(gè)看家基因表達(dá)，更好定量靶標(biāo)基因的水平。
這類引物系列包括人、小鼠和大鼠三個(gè)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(shí)的引物序列，用于研究不同成員的表達(dá)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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