...(~45 minutes). Method #2 Place mixed oligos in a PCR tube. Place tube in a thermocycler programmed to ...

...commonly used technique for molecular cloning, such as PCR- or restriction enzyme-based cloning. Purifying DNA...maintaining sterility while working. AAV Titration by qPCR Use qPCR to measure the physical titer of your adeno-...adeno-associated virus preparation. AAV Titration by qPCR Using SYBR Green Technology Protocol Agarose Gel...

... This is frequently done after performing either PCR - or restriction enzyme -based cloning to test individual...

...commonly used in molecular cloning techniques, such as PCR or restriction cloning . It is also used to quickly...

...and insert DNA to use. Combine the following in a PCR or Eppendorf tube: Vector DNA Insert DNA Ligase Buffer...

...minutes at 95°C in a PCR machine or in a beaker of boiling water. If using a PCR machine, incubate the...Days post-infection mRNA knockdown (quantitative PCR) ≥3 days Protein knockdown (western blot) ≥4 days...

... onto the insert of interest using PCR. This is accomplished by PCR amplifying the insert sequence using...restriction sites onto the insert of interest using PCR. The process of subcloning consists of digesting ...

...restriction sites to flank your insert : You can use PCR Based Cloning and add restriction sites to the ends...

...surviving cells are collected, their plasmids are PCR-amplified and sequenced using NGS to identify their...

...controlling GPCR activation have been developed to selectively study individual types of GPCRs with greater...be used for lower frequency stimulation. GPCR signaling GPCR signaling is a complicated process involving...change the membrane potential. OptoXRs are modified GPCRs. When exposed to light, they activate downstream... coupled receptors  G-protein coupled receptors (GPCR) regulate cellular function through multiple intra-cellular...have been developed for utilizing light to control GPCR activation and downstream signaling cascades (reviewed...Levitz, 2020), including light-sensitive chimeric GPCRS (OptoXRs) (Tichy et al., 2019), photoswitchable ...rhodopsin, and the intracellular domain of an endogenous GPCR, such as adrenergic receptors. When exposed to light...

...requires ddPCR skills. Your resume lists ddPCR, so the hiring manager knows that you have ddPCR experience...with ddPCR, but do have experience with qPCR, you could talk about how your experience with qPCR would...that by describing how you developed and used a ddPCR assay to answer an experimental question. Now, the...manager knows that you can design experiments with ddPCR, interpret the results, and develop a successful...would make it easy to master ddPCR.  Finally, it’s a good idea to state why you’re applying to the specific...

...biosensors kit By Mike Lacy G protein-coupled receptors (GPCRs) are critical components of many physiological pathways...targets. To enable large-scale profiling of many GPCRs, Mikel Garcia-Marcos’ lab recently developed a collection... detection of activation triggered by endogenous GPCR. Example applications include large-scale pharmacological...The authors used this platform to assay dozens of GPCRs across many cell types in varying conditions, revealing...2024). Direct interrogation of context-dependent GPCR activity with a universal biosensor platform. Cell...

...titer results between ddPCR and qPCR? A: At CLOVER, we have traditionally used qPCR (protocol link). We ...this to strongly bias titer. We are also adopting ddPCR, which does not a standard curve and somewhat more...

...titration by qPCR using SYBR green technology Here’s some advice on how to successfully set up your qPCR to determine...generate a standard curve, visit the AAV Titration by qPCR written protocol page. Getting started with tissue...

...single-stranded DNA sequence used as an initiation point for PCR amplification or DNA sequencing of the plasmid. Primers...

...signaling pathways G protein-coupled receptors (GPCRs) detect molecules outside of the cell and translate...those signals into cellular responses. For example, GPCRs and their downstream G protein partners can regulate...have a long standing interest in the biology of GPCRs and G proteins, with an emphasis on oligomerization...

...clinical and diagnostic labs, deep sequencing and qPCR methods are too costly and time intensive to process...platform on clinical stool samples by comparison to qPCR and demonstrated that it can also detect expression...mRNA expression cannot be identified with DNA-based qPCR methods, the ability to detect mRNA transcripts ...

...designer drugs, are G protein-coupled receptors (GPCRs), that have been reprogrammed to respond only to... threshold and silence neurons. DREADD-activated GPCR signaling can also modify other properties such ...

... diagnosing COVID-19, aside from the widely used qPCR and antigen tests.  Labs including Jennifer Doudna...Taq polymerase that can be used in a one-step RT-qPCR reaction to detect SARS-CoV-2 nucleic acids. Mammalian...

... loop regions of any G-protein-coupled receptor (GPCR) recognizing a specific ligand (Ino et al. 2022)...produce more than 20 other MTRIA sensors based on GPCRs recognizing various neurotransmitters, neuropeptides...

...promoters (orange). Hybridoma-derived V L and V H domain PCR products are fused to a joining fragment comprising...translocation of the heavy chain across the ER membrane. PCR-mediated fusion of these three elements is followed...

...pBS500 EF1alpha-GFPcre Cre-GFP fusion EF-1 alpha Mammalian Sauer 11923 pBS598 EF1alpha-EGFPcre Cre-EGFP fusion...EF1alpha-EGFPcre* Cre-EGFP fusion EF-1 alpha Mammalian Sauer 11956 pBS594 promoterless EGFPcre Cre-EGFP...tet-hCMV-EGFPcre Cre-EGFP fusion Tet inducible Mammalian Sauer 11958 pBS532 promoterless GFPcre promoterless...-hCMV-GFPcre tet inducible Cre-GFP fusion tet-hCMV Mammalian Sauer 11961 pBS596 tet-hCMV-GFPcre tet inducible...Cre EF-1 alpha Mammalian Sauer 11919 pBS448 RSV-GFPcre Cre-GFP fusion Rous sarcoma virus (RSV) LTR/promoter...Tamoxifen-inducible Cre None Zebrafish Stankunas 82696 pCRE-iRFP670 Cre and iRFP670 PGK Mammalian Mullen 84032...

...PROCR protein C receptor, endothelial (EPCR) CCCA, CCD41, CD201, EPCR, MGC23024, bA42O4.2 PSMB8 proteasome...receptor 2 CYSLT2, CYSLT2R, GPCR, HG57, HPN321, KPG_011, PSEC0146, hGPCR21 DARC Duffy blood group, chemokine...relaxin/insulin-like family peptide receptor 3 GPCR135, MGC141998, MGC142000, RLN3R1, RXFPR3, SALPR SAA1... estrogen receptor 1 CEPR, CMKRL2, DRY12, FEG-1, GPCR-Br, GPR30, LERGU, LERGU2, LyGPR, MGC99678 GPHA2 ...repeat-containing G protein-coupled receptor 6 FLJ14471, GPCR, VTS20631 LHB luteinizing hormone beta polypeptide...relaxin/insulin-like family peptide receptor 3 GPCR135, MGC141998, MGC142000, RLN3R1, RXFPR3, SALPR RXRA...

...quality control processes as our in-stock service: ddPCR, purity testing, and sterility testing, plus viral...

...new optogenetic tool called PdCO.  This is an "optoGPCR" (coupled to Go, so it's an inhibitory opsin!)...

...assays we run with all of our in-stock viral preps: ddPCR, purity testing, sterility testing, and viral genomic...

...advantages Mapping the brainDissecting circuits Studying GPCR pathways   Acknowledgements We thank the Bryan...

.... Used for photoswitchable control of inhibitory GPCR signaling cascades. Michael Bruchas 139505-AAV8 .... Used for photoswitchable control of inhibitory GPCR signaling cascades. Michael Bruchas 139505-AAV9 .... Used for photoswitchable control of inhibitory GPCR signaling cascades. Michael Bruchas 140135-AAV1 ...

...for downstream applications (Berns, 1972). Using qPCR and restriction enzyme based methods, we showed ...

...for the newest GRAB sensors from the Li Lab. These GPCR based sensors provide the spatiotemporal resolution...

... in gene expression readouts from assays such as qPCR or microarray. But, aside from facilitating the ...

... gRNAs can be expressed in a single vector, pCRISPRainbow-DONOR1, with dCas9 supplied by a separate vector...

...Rhodopsin GEVIs Rhodopsin is composed of an opsin (a GPCR) linked to a chromophore and functions as a photoreceptor...

...viral vector production facilities frequently use qPCR with a standard curve to titer their viruses; food...

...cells.  While Addgene uses a physical titer (via ddPCR) to quality control our viral preps, we recommend...

... of the new viral preps from recent months: New GPCR activation-based norepinephrine (GRABNE) sensors...

...laboratories. In the case of the COVID-19 pandemic, qPCR can be used to diagnose the presence of SARS-CoV...

...based on a split NanoLuc® have been used to assess GPCR-G protein interactions , protein dimerization , ... firefly luciferase and GFP James Wilson 106457 pCR3.1-Luc Firefly CMV Mammalian expression of firefly...

...and the reverse transcriptase activity assay uses qPCR to quantify viral RNA. All of these methods have...

...Parkinson's Mark Cookson 29354 pCR8-DJ1-WT PARK7 Parkinson's Mark Cookson 29355 pCR8-DJ1-D149A PARK7 Parkinson's...Parkinson's Mark Cookson 29356 pCR8-DJ1-E64D PARK7 Parkinson's Mark Cookson 29357 pCR8-DJ1-M26I PARK7 Parkinson's...-LFA EIF2AK2 GAL VWM disease Harmit Malik 20123 PCRII-zSynC SNCB Lewy body dementia Aaron Gitler 20648...SOD1 GFP CMV ALS Elizabeth Fisher 26573 CaV2.1 (pCR3.1) CACNA1A CMV Spinocerebellar ataxia 8 Diane Lipscombe...

...key, especially for G-protein-coupled receptors (GPCRs). Mutations in receptor sites can affect the fluorescence...

...used either with plasma membrane proteins (such as GPCRs, ion channels, or cell surface receptors) or with...

...RNAi sequences, confirmation of RNA knockdown by RT-qPCR, and potentially the need to study protein expression...

...eabq8173. Yulong Li Norepinephrine nLight red and green GPCR-based NE indicators Sensitive multicolor indicators...Tommaso Patriarchi Norepinephrine GRAB_NE family of GPCR activation-based NE sensors A Genetically Encoded...

... is necessary for Cas enzymes to bind target DNA PCR P olymerase C hain R eaction; Used to amplify a specific...

...cCGCTTCAGCggtctGTGGACTCGa zebrafish similar to grp151 (or gpcr-2037) OZ535 and OZ536 OPEN OPEN aGCCATCATCaggttcaGGATGAGCCc...

...three-part fusion proteins - Planas and Santalucia Labs pCri System : Heterologous protein expression in E. coli...

...lab Cas9 species = S. pyogenes (PAM = NGG) 42875 pCRISPR BsaI E. coli, S. pneumoniae Kanamycin none, need...

...undergo quality control, including AAV titration by ddPCR, in vitro and (when possible) in vivo testing, and...

...Q157TAG Plasmid 53225 MBP-MEK1 S218TAG/S222TAG ( aka. PCRT7 tetR pLtetO MBP-MEK1 XX Amp) Plasmid 68300 MBP-MEK1...