Skip to main content
Addgene

Enter a sequence to perform a BLAST-based search of the Addgene plasmid sequence database.

Information

In Category

We narrowed to 400 results for: pho;

Showing: 351 - 400 of 400 results

  2. Site Directed Mutagenesis by PCR

    Type
    Blog Post
    ...plasmids for the mutation of interest. Gel electrophoresis diagrams of the anticipated restriction analysis...

  11. Affinity Purification of Recombinant Antibodies with Protein A or Protein G

    Type
    Protocol
    ...431098 Sodium phosphate monobasic monohydrate, Millipore Sigma 71507-250G Sodium phosphate dibasic, Millipore...sterilize 1 M of sodium phosphate buffer, pH 7.0 390 mL of sterile sodium phosphate monobasic monohydrate...binding buffer (20 mM sodium phosphate buffer, pH 7.0) 20 mL 1M sodium phosphate buffer 980 mL of deionized...compatible with 50 mL conical tubes NanoDrop spectrophotometer 37 °C, 5% CO 2 incubator with shaking platform...RT) before use. Reagent Preparation 1 M sodium phosphate monobasic monohydrate (NaH 2 PO 4 ∙H 2 O), pH ... 7.0 Autoclave or filter sterilize 1 M sodium phosphate dibasic (NaH 2 PO 4 ), pH 7.0 142 g of NaH 2 PO... (NaH 2 PO 4 ∙H 2 O) 610 mL of sterile sodium phosphate dibasic (NaH 2 PO 4 ) Store up to 1 month at 4...

  12. Protocol - How to Ligate Plasmid DNA

    Type
    Protocol
    ...with a phosphatase before performing the ligation reaction (phosphatase removes the 5' phosphate and therefore...was completely digested and if phosphatase treated, that the phosphatase treatment worked. This control... ligase catalyzes the formation of covalent phosphodiester linkages, which permanently join the nucleotides... vector re-circularization - most useful for phosphatase treated vector Insert or water + Any colonies...

  13. DNA Quantification

    Type
    Protocol
    ...purity. This can be done with a spectrophotometer. A spectrophotometer uses the absorbance/transmission... instructions for the spectrophotometer specific to your lab. Spectrophotometer Tips Before measuring ...labs have a NanoDrop, which is a very small spectrophotometer that can accurately read DNA concentration...measuring any samples, be sure to ‘blank’ the spectrophotometer using the solution the DNA is resuspended in, ...NanoDrop. It is a good idea to re-zero any spectrophotometer between samples if they are expected to vary...

  14. Protocol - How to Run an Agarose Gel

    Type
    Protocol
    ... Protocols Agarose Gel Electrophoresis Agarose Gel Electrophoresis You may also like... Restriction...Standard protocol for performing agarose gel electrophoresis, including tips to improve resolution and separation... Agarose Gel DNA Ligation Introduction Gel electrophoresis is the standard lab procedure for separating...pairs) for visualization and purification. Electrophoresis uses an electrical field to move the negatively...protocol video below to learn how to perform gel electrophoresis. Equipment Casting tray Well combs Voltage ...solidified, place the agarose gel into the gel box (electrophoresis unit). Fill gel box with 1xTAE (or TBE) until... short DNA fragments is polyacrylamide gel electrophoresis (PAGE), which is typically used to separate...

  15. Coomassie Purity Stain of Recombinant Antibodies

    Type
    Protocol
    ...separated by denaturing polyacrylamide gel electrophoresis alongside serial dilutions of a standard antibody...pipette Pipette controller Microcentrifuge Electrophoresis chamber Power supply Rocking platform Fume ...the bottom of the gel. Place the gel in the electrophoresis chamber and secure it. Note: Check the manufacturer...make quantifying the gel easier. Cover the electrophoresis chamber and attach to a power supply. Run the.... Turn off the power supply and unplug the electrophoresis chamber. Remove the gel from the chamber. Use...background staining. Pro-Tip A large shift in the electrophoretic mobility of the bands (sample AR0016 in Figure...ImageJ (Link opens in a new window) or a similar photo software to determine the relative intensity of ...

  16. Molecular Biology Protocol - Restriction Digest of Plasmid DNA

    Type
    Protocol
    ...insert. CIP (calf alkaline phosphatase) or SAP (shrimp alkaline phosphatase) are commonly used. Follow...your restriction digest results. Equipment Electrophoresis chamber Pipetman Reagents Liquid DNA aliquot...manufacturer's instructions) Gel loading dye Electrophoresis buffer Pipet tips Procedure Select restriction...visualize the results of your digest, conduct gel electrophoresis . Tips and FAQ Restriction enzymes MUST be ...enzyme to cut the vector, you will need to use a phosphatase to prevent re-circularization of the vector if...

  17. Plasmid Cloning by Restriction Enzyme Digest (with Protocols)

    Type
    Protocol
    ... the phosphatase you choose. CIP (calf alkaline phosphatase) or SAP (shrimp alkaline phosphatase) are ...to use a single enzyme, but this will require phosphatase treatment of your recipient plasmid as well as...overhangs after digestion, you will need to use a phosphatase to prevent re-circularization of the recipient...treat your digested recipient plasmid with a phosphatase prior to the ligation step or prior to the gel...

  18. Protocol - How to Perform a Diagnostic Digest

    Type
    Protocol
    ...Restriction Digest of Plasmid DNA Agarose Gel Electrophoresis Introduction Restriction enzymes are naturally...and analyze the resulting fragments by gel electrophoresis . The pattern of the fragments on the gel can...to analyze a restriction digest. Equipment Electrophoresis chamber Pipetman Pipet tips Reagents Liquid...manufacturer's instructions) Gel loading dye Electrophoresis buffer Verifying Total Plasmid Size -OR- Insert... tricks see the Restriction Digest and Gel Electrophoresis pages....

  19. Plasmid Cloning by PCR (with Protocols)

    Type
    Protocol
    ... the phosphatase you choose. CIP (calf alkaline phosphatase) or SAP (shrimp alkaline phosphatase) are ...overhangs after digestion, you will need to use a phosphatase to prevent re-circularization of the vector. ... treat your digested recipient vector with a phosphatase prior to the ligation step or prior to the gel...

  20. Protocol - How to Purify DNA from an Agarose Gel

    Type
    Protocol
    ...gel electrophoresis, which separates DNA by their length in base pairs. Following electrophoresis, you...Restriction Digest of Plasmid DNA Agarose Gel Electrophoresis DNA Ligation Background Information Gel purification...Protocol: Gel Purification Follow the Agarose Gel Electrophoresis Protocol with the following amendments: Notes...

  21. Video Library

    Type
    Protocol
    ... Technology Protocol Agarose Gel Electrophoresis Gel electrophoresis is the standard laboratory procedure...visualization and purification. Agarose Gel Electrophoresis Protocol Purifying DNA from an Agarose Gel ...DNA fragments based on size. Following gel electrophoresis, you can cut DNA bands out of the agarose gel...

  22. General Transfection

    Type
    Protocol
    ...min. 25 mM chloroquine diphosphate Dissolve 0.129 g of chloroquine diphosphate salt into 10 mL of sterile...fresh DMEM complete containing 25 µM chloroquine diphosphate and incubate ~5 h For 10 mL of DMEM complete,...complete, add 10 µL of 25 mM chloroquine diphosphate. Dilute 18.9 µg of DNA into 500 µL of Opti-Pro SFM. Pro-...

  24. Pipetting Protocol

    Type
    Protocol
    ...Information Anatomy of a Pipette This front view photo of a P1000 pipette shows the plunger at the top,... pipette tip off of the pipette. This side view photo of a P1000 pipette shows the volume adjustment ring...ready to begin using the pipette. To the right is a photo of how you might hold a pipette. Without moving ...

  25. Antibody Validation Using the Indirect ELISA Method

    Type
    Protocol
    ...of each well at 450 nm on the spectrophotometer. Use the spectrophotometer to generate a standard curve...an automated ELISA plate washer. Equipment Spectrophotometer compatible with 96-well plates 1–10 µL single...

  26. Ligation Independent Cloning

    Type
    Protocol
    ...cloning scars"). A “stuffer” sequence allows for electrophoretic separation of linearized vector from the reaction...linearized vector from the reaction mixture by gel electrophoresis followed by gel purification . The cut vector...

  28. What is Polymerase Chain Reaction (PCR)

    Type
    Protocol
    ...product can be verified based on size by gel electrophoresis . Protocol Video Watch the protocol video below...rate of the DNA polymerase. Deoxynucleoside Triphosphates (dNTPs): Are the building blocks added one at...

  29. Kit Free RNA Extraction

    Type
    Protocol
    ...the quality of your RNA sample(s) using a spectrophotometer (such as a Nanodrop), agarose gel, or bioanalyzer...

  31. Protocol - How to Design Primers

    Type
    Protocol
    ...Reaction Plasmid Cloning by PCR Agarose Gel Electrophoresis Primer Design for PCR Oligonucleotide primers...

  32. Weighing Reagents Protocol

    Type
    Protocol
    ...will be weighed out Procedure The balance in the photo above has a capacity of 200 g, as indicated by the...

  33. Centrifugation

    Type
    Protocol
    ... will have essentially the same components. The photo to the left shows a front view of an open tabletop...

  36. Western Blot

    Type
    Protocol
    ...Pipette controller Pipette tips and pipettes Spectrophotometer Heat block Mini gel tank chamber Power supply...

  38. Lentivirus ddPCR Titration

    Type
    Protocol
    ...Determine the concentration of each sample on a spectrophotometer. Prepare 25 ng/µL stocks of each sample. Samples...

  39. Optogenetics Guide

    Type
    Guide
    ...)/~500 (off) PhoCl Photocleavable protein engineered from a green-to-red photoconvertible fluorescent ...enhancements include: Increased photocurrent amplitude Examples: iChloC, SwiChRca, Phobos, Aurora Browse Channelrhodospin...reduces photocurrent amplitude 490 ChR/T159C T159C mutation; displays increased photocurrents 470 SFO/...bacterial photoreceptors to create protein systems controlled by light. These “photoswitchable” proteins...Kv) channels fused to LOV2 photoswitch. Illumination causes the photoswitch to unfold, lowering the toxin's...synthetic, photosensitive kinase, which uses the ubiquitous flavin mononucleotide as its chromophore, and phosphorylates... 400 UVR8 domains UVR8 plant photoreceptor protein that forms photolabile homodimers with slow reversal...

  40. Antibody Guide

    Type
    Guide
    ... curve with known amounts of protein. Fluorophores Fluorophores are light-sensitive molecules which emit...IF uses fluorophore color to differentiate between the targets of interest, if your fluorophore is bound...the fluorophores used is important. It is best to inversely pair the brightness of the fluorophore with...If a bright fluorophore is on an abundant target, it obscures fainter signals. Fluorophores can emit signals...storing your antibody: Antibodies conjugated to fluorophores should never be stored below 4 °C. Certain isotype...molecules are horseradish peroxidase (HRP) and fluorophores. Horseradish peroxidase (HRP) HRP performs a...specific wavelength. While more expensive than HRP, fluorophores are available in a range of colors activated...

  41. Gamma-Retroviral Vector Guide

    Type
    Guide
    ...envelope, Phoenix-ECO (for infection of mouse and rat cells), or an amphotropic envelope, Phoenix-AMPHO (for...of the viral packaging cell line. For example, Phoenix™ is a popular second-generation gamma-retroviral... produced in just a few days. Another variant, Phoenix-gp, contains only gag-pol and enables additional...

  42. Chemogenetics Guide

    Type
    Guide
    ...activate neuronal firing through stimulating phospholipase C, which releases intracellular calcium stores...molecule driven by FRET-modified bioluminescence. Neurophotonics. 11:021005. PMID: 38450294 (Link opens in a...luminopsins with improved coupling efficiencies. Neurophotonics. 11:024208. PMID: 38559366 (Link opens in a...

  43. CRISPR Guide

    Type
    Guide
    ...SpCas9-HF1 - disrupt Cas9’s interactions with DNA phosphate backbone HypaCas9 - increase Cas9 proofreading...fusing the anti-CRISPR protein AcrIIA4 with the photosensitive LOV2 domain . Several labs have also controlled...: Epigenetics Visualizing Genomic Loci Using Fluorophores CRISPR imaging has numerous advantages, including...11: Overview of visualizing genomic loci with fluorophores using CRISPR Browse Plasmids: Visualize Purify...Yoshimi, K., Okuzaki, Y., Gee, P., Kunihiro, Y., Sonpho, E., Xu, H., Sasakawa, N., Naito, Y., Nakada, S...

  44. Molecular Biology Reference

    Type
    Guide
    ...of restriction enzymes, DNA ligase, and gel electrophoresis allowed for the ability to move specific fragments...Delta(lacZYA-argF) U169 recA1 endA1 hsdR17(rk-, mk+) phoA supE44 thi-1 gyrA96 relA1 tonA JM109 Addgene; Promega...4 standard bases are tagged with a different fluorophore so they can be distinguished from one another...

  45. Promoters

    Type
    Guide
    ...promoter PGK Constitutive Mammalian promoter from phospholycerate kinase gene TRE Inducible Tetracycline response...

  46. Plan Your Experiment

    Type
    Guide
    ...gRNA Presence of a selectable marker (drug or fluorophore) Delivery method Expression System Components...identify DNA fragments. PCR amplification and gel electrophoresis (for HDR or NHEJ): For large deletions or insertions...

  47. Educational Resources

    Type
    Guide
    ...plasmid cloning, and virus, including: Gel Electrophoresis Bacterial Transformation Plasmid Cloning Creating...

  48. Addgene's eBook Collection

    Type
    Guide
    ...localization; an introduction to non-protein fluorophores; and much more! Close About Overlay Select Viral...

  49. Lentiviral Vector Guide

    Type
    Guide
    ... are other envelope gene options, including: Lymphocytic Choriomeningitis Virus (LCMV) – less toxic and...
Showing: 351 - 400 of 400 results