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We narrowed to 802 results for: mak

Showing: 51 - 100 of 802 results

  1. Chemogenetics AAV Preps

    Type
    Collection
    ... vast catalog of eligible AAV plasmids and we’ll make the viral vector prep you need. Or fill out our ...

  2. Antibody Plasmid Collection

    Type
    Collection
    ...102. Joanna Bereta Sybody Generation Toolbox for making synthetic nanobodies (sybodies) against membrane...

  3. Microbiology Resources

    Type
    Collection
    ...Engineering blog post to see how Addgene depositors are making this possible in a range of organisms. Find CRISPR...

  4. Worm Expression Resources

    Type
    Collection
    ...hermaphrodite consists of 959 somatic cells, which makes them attractive for imaging and fluorescence microscopy...

  5. Zebrafish Plasmid Collection

    Type
    Collection
    ...nucleases, TALENS, and CRISPR/Cas9-based gene editing make the zebrafish a stalwart research model. With more...

  6. Control AAV Preps

    Type
    Collection
    ... vast catalog of eligible AAV plasmids and we’ll make the viral vector prep you need. Or fill out our ...

  8. Lentivirus Plasmids

    Type
    Collection
    ...components Trono 63586 psPAX2-D64V 2nd Packaging For making Integrase deficient lentivirus Rawlings , Scharenberg...

  10. COVID-19 Resources

    Type
    Collection
    ...and Yeast expression Spike protein constructs for making VSV pseudo virus Mammalian constructs Due to the...

  11. Optogenetics AAV Preps

    Type
    Collection
    ... vast catalog of eligible AAV plasmids and we’ll make the viral vector prep you need. Or fill out our ...

  12. Retrograde AAV viral preps

    Type
    Collection
    ... vast catalog of eligible AAV plasmids and we’ll make the viral vector prep you need. Or fill out our ...

  13. Caltech Systemic Capsids

    Type
    Collection
    ... vast catalog of eligible AAV plasmids and we’ll make the viral vector prep you need. Or fill out our ...

  14. Tetracycline Inducible Expression

    Type
    Collection
    ...conditional auxin-inducible degron system Masato Kanemaki 92099 AAVS1_Puro_Tet3G_3xFLAG_Twin_Strep Tet-inducible...

  15. Biosensor AAV Preps

    Type
    Collection
    ... vast catalog of eligible AAV plasmids and we’ll make the viral vector prep you need. Or fill out our ...

  17. CRISPR Guide

    Type
    Guide
    ...generating gRNAs makes CRISPR one of the most scalable genome editing technologies, making CRISPR perfect... Gootenberg, J. S., Abudayyeh, O. O., Slaymaker, I. M., Makarova, K. S., Essletzbichler, P., Volz, S. ...very close proximity to the region to be edited, making the PAM sequence (NGG) limiting. This PAM is abundant...Anti-CRISPR Small Precision Edits CRISPR can be used to make small, very precise edits of a handful of bases,...RNA-binding protein to stabilize pegRNAs Twin PE - two PEs make complementary edits to opposite strands Prime editing...the ability to target almost any genomic locus, makes CRISPR an ideal genome engineering system for large-scale...large-scale, forward genetic screening. CRISPR can make highly specific, permanent genetic modifications...

  18. Pouring LB Agar Plates

    Type
    Protocol
    ...pouring, you should stop pouring and re-make the gel-mix. If you’re making plates without any antibiotic you...resistance gene. The following protocol will allow you to make your own LB/agar plates with your antibiotic of ...-agar powder per L of molten agar you’d like to make. The precise mass you measure out will be based ... like to pour. For example: Because we’d like to make 20 plates, and our plates can hold a maximum of ...200 mL of media total. Importantly, we’ll actually make a bit more than 200 mL (~220 mL) just in case we...small errors in measurement. You should also always make a bit more gel-ager mix than you think you’ll need...an appropriately sized bottle for autoclaving. We make 400 mL of agar in 1 L bottles and 200 mL of agar...

  19. Molecular Biology Reference

    Type
    Guide
    ...these 4 nucleotides makes up the genetic code and provides the instructions to make every protein within...these bases makes up the genetic code and provides all the information needed for cells to make proteins ...Once you have constructed a plasmid, you can easily make an endless number of copies of the plasmid using... can be dissolved in dH 2 0. Addgene recommends making 1000X stock solutions and storing aliquots at -...into your LB medium at 1:1,000. For example, to make 100 mL of LB/ampicillin growth media, add 100 μL...of the known primer region with very few errors making it an efficient and reliable sequencing method....A,C,T, or G) is labelled with a different color making it easy to identify the order of the DNA strand...

  20. AAV Titration by qPCR Using SYBR Green Technology

    Type
    Protocol
    ... such as pipetting error and sample evaporation. Make the master mix after all the samples have been added...Pro-Tip Once a validated standard curve is obtained, make a small aliquot of each standard (enough for 1 or...value of the standard starts to drift, it’s time to make a new one. When developing the assay multiple plasmids...do NOT treat your plasmid standard with DNase ** Make 6 serial dilutions, in duplicate, of your standard...quality of the sample dilution series is critical. Make sure to pipet each dilution up and down at least...standard curve and the sample dilutions. Pro-Tip Make sure that the qPCR is valid by checking to the following...you should observe differences in Ct values that make sense for your dilutions (~3.3 difference Ct for...

  21. Weighing Reagents Protocol

    Type
    Protocol
    ...you’ll need to make buffers, media, or other solutions. A key part of this task is making sure you’re weighing...weighing by looking for a weight range on the scale. Make sure that the weight of the material you’re weighing...add the material directly to the tube to weigh it. Make sure that your scoopula or spatula is clean, and...of your reagent, transfer it into your container. Make sure that any spills produced during the protocol...you have your reagents weighed out, you can begin making your solutions!...

  22. Isolating a Monoclonal Cell Population by Limiting Dilution

    Type
    Protocol
    ... growth. This conditioned medium will be used to make a cell solution later in this protocol. Isolating...solution into the conditioned medium prepared above to make a new cell solution at a concentration of 5 cells...concentration: 4 × 10 5 cells/mL To seed one 96-well plate, make 10 mL of a 5 cell/mL solution. Calculate the total...0.125 µL Because this is such a small volume, first make 1 mL of a 1:100 dilution of the homogenized cell...transfer 100 times that volume, which is 12.5 µL. To make the final 5 cell/mL solution, transfer 12.5 µL of... the same focal plane. Scan the entire plate and make note of each well in which you see growth. At this...

  23. Lentiviral Vector Guide

    Type
    Guide
    ...engineered lentiviral vectors have many aspects that make them useful research tools in a variety of cell ...plasmids also contain a deletion in the 3’ LTR to make them self-inactivating (SIN). This deletion is transferred... long-term, constitutive expression. In order to make stable cell lines, VSV-G is often put under an inducible...Stable Cell Lines Lentiviral vectors can be used to make stable cell lines due to their integration into ... by daughter cells. Inheritance of the barcodes makes lentiviral libraries popular for lineage tracing...

  24. Adeno-associated virus (AAV) Guide

    Type
    Guide
    ...number will continue to increase. This diversity makes rAAV vectors a very useful system for preferentially...capacity for long-term gene expression. These features make them ideal for gene therapy, neuroscience research...channelrhodopsins, halorhodopsins, and other optogenetic genes makes them suitable to be packaged in the relatively small...choice for the delivery of CRISPR/Cas9 elements, making it one of the most common methods for in vivo CRISPR-based...shown higher efficiency in delivering large genes makes use of split inteins, proteins that carry out a ...

  25. Protocol - How to Run an Agarose Gel

    Type
    Protocol
    ...volume to make gels of other agarose concentrations (e.g., 2 g of agarose in 100 mL of TAE will make a 2% ...is very little difference between the two. Note: Make sure to use the same buffer as the one in the gel... Rule For each sample you want to load on a gel, make 10% more volume than needed because several microliters...For example, if you want to load 1.0 μg in 10 μL, make 1.1 μg in 11 μL. Reference Page Top Index...

  26. Protocol - How to Create a Bacterial Glycerol Stock

    Type
    Protocol
    ...stocks of their plasmids. This way, when you want to make more plasmid DNA, the plasmid will already be in...screw top tube or cryovial and gently mix. Notes: Make the 50% glycerol solution by diluting 100% glycerol...liquid bacterial culture, take 500 μL of culture to make your glycerol stock before you begin your plasmid... shake the glycerol before freezing (5-6 times). Make sure that you see one uniform solution, and there...

  27. Lab Safety for Biosafety Levels One and Two

    Type
    Protocol
    ...chew gum, or apply makeup in the laboratory. Before you start your experiment, make sure your workspace...and the appropriate contact times for each agent. Make sure you have enough space to work at your lab bench...extinguisher are also required to be present in the room. Make sure that you know where these are located before...

  28. What is Polymerase Chain Reaction (PCR)

    Type
    Protocol
    ...38.5nm, add 385µl of water. After making your 100uM stock, immediately make a working concentration of each...their primers in liquid, normally sterile dH 2 O. To make a 100uM stock of any primer, add a number of µl ...each primer (10uM) by making a 1:10 dilution of the stock. For example, add 100µl of primer stock to 900µl...

  29. Kit Free RNA Extraction

    Type
    Protocol
    ...precipitation step, Option B) Glycogen (Optional) Caution Make sure to read the SDS (Safety Data Sheet) for safety...Solution D Protocol Before starting this protocol, make sure to prepare solution D (see reagent section ...freeze-thaw cycles of your entire RNA sample, consider making smaller aliquots of it and storing those in -80...

  30. Guide to Using Pooled Libraries

    Type
    Guide
    ...plasmid libraries, the plasmids must first be used to make virus. This pooled virus is subsequently used to...that an infected cell receives only one plasmid. To make sure that every plasmid is adequately represented...electroporation and maxiprep). If delivering as virus, make virus; this creates a pooled lentiviral CRISPR library...

  31. Plan Your Experiment

    Type
    Guide
    ...reagents are appropriate for a given experiment. Make sure to check whether reagents are available to ...efficiency compared to NHEJ knockout. Base editors can make a limited set of mutations. Repress or Interfere...envelope plasmids provide the necessary components to make lentiviral particles (for details about lentivirus...

  32. Chemogenetics Guide

    Type
    Guide
    ...exhibited high levels of constitutive activity, making them less than ideal. DREADDs Designer Receptors... previously used in human populations in Japan, making it an attractive option for translational studies... workflows (such as laser or LED illumination), making LMOs a versatile option. Coupling brighter bioluminescent...

  33. Optogenetics Guide

    Type
    Guide
    ... invasive. Different activation wavelengths also make it possible to combine multiple opsins in the same...196: 1-28. PMID 22341318 Gradinaru V, Zhang F, Ramakrishnan C, Mattis J, Prakash R, Diester I, Goshen I,...PMID 20621963 Mattis J, Tye KM, Ferenczi EA, Ramakrishnan C, O'Shea DJ, Prakash R, Gunaydin LA, Hyun M...

  34. Gamma-Retroviral Vector Guide

    Type
    Guide
    ... gamma-retroviral vectors have many aspects that make them useful research tools in a variety of cell ... long-term, constitutive expression. In order to make stable cell lines, VSV-G is often put under an inducible...Lines Gamma-retroviral vectors are often used to make stable cell lines. Like their popular lentiviral...

  35. Protocol - How to Streak a Plate

    Type
    Protocol
    ...Bacteria on an LB Agar Plate You may also like... Making LB Agar Plates Bacterial Transformation Recovering...the way you would hold a pencil, so that you can make a broad stroke. Only touch the surface of the plate... diagonal lines in another section of the plate. Make sure that the first line (and only the first) in...

  36. Gibson Assembly Protocol

    Type
    Protocol
    ... with a two-part Gibson reaction if you're only making a small change in a plasmid (such as point mutations...manufacturer's instructions. You can purchase master mix or make your own. Transform bacteria with the DNA and screen...overlapping PCR primers (>60 bp) but too short to make its own part (<150 bp). Pro-Tip Please note that...

  37. Using a Light Microscope Protocol

    Type
    Protocol
    ...of a microscope are magnification (the ability to make an image larger) and resolution (the ability to ...through the sample. However, some of that light won’t make it through the sample because different components...fine focus knob (the smaller of the focus knobs) to make minor adjustments to the focus. After you are happy...

  38. Antibody Guide

    Type
    Guide
    ...plasmids. Animals - Antibodies produced in animals make use of the animal’s immune system. The antigen of...Overview of application Collect and lyse samples to make proteins available. Tissue samples may need additional...Ready to put your new knowledge to use? Addgene now makes ready-to-use recombinant antibodies made from plasmids...

  39. AAV Purification by Iodixanol Gradient Ultracentrifugation

    Type
    Protocol
    ... of the rotor and place them in a stable rack. **Make sure not to disturb the gradient!** Collect Fractions...flow from the needle set at the 40–60% interface. Make sure that the microcentrifuge tubes are well positioned...spin, add more formulation buffer and sample and make sure to pipet back and forth a few times to mix ...

  40. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ... mouse genes. Addgene is working with the TRC to make this shRNA cloning vector available to the scientific... microfuge tubes (do NOT use polystyrene tubes), make a cocktail for each transfection: 1 μg pLKO.1 shRNA...scientists with high-quality goods and services. Addgene makes every effort to ensure the accuracy of its literature...typographical or other errors may occur. Addgene makes no warranty of any kind regarding the contents of...

  41. AAV ddPCR Titration

    Type
    Protocol
    ...the risk of contaminating reagents we recommend making small aliquots of master mixes, primers, and probes...and place in the Biological Safety Cabinet (BSC). Make sure that the BSC to be used for dilution and the...wells. Cover the cartridge with the DG8 gasket, making sure that it is secure. Transfer the cartridge ...

  42. Lentivirus ddPCR Titration

    Type
    Protocol
    ...place it in the Biological Safety Cabinet (BSC). Make sure that the BSC is supplied with sufficient pipette...wells. Cover the cartridge with the DG8 gasket, making sure that it is secure. Transfer the cartridge ...the risk of contaminating reagents we recommend making small aliquots of master mixes, and primers/probe...

  43. Cloning

    Type
    Guide
    ...repeated throughout a specific DNA sequence, which can make it difficult at times to identify compatible restriction...site-specific recombination or a ligation step, making it an easy, cheap and rapid cloning method. So ...

  44. Protocol - Over-Agar Antibiotic Plating

    Type
    Protocol
    ...Over-Agar Antibiotic Plating You may also like... Making LB Agar Plates Bacterial Transformation Recovering...transformed E. coli . Over-agar spreading of antibiotic makes it easy for an investigator to conveniently plate...

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

    Type
    Protocol
    ... Rule For each sample you want to load on a gel, make 10% more volume than needed because several microliters...For example, if you want to load 1.0 μg in 10μL, make 1.1μg in 11μL. Reference Page Top Index...

  46. Protocol - How to Perform Sequence Analysis

    Type
    Protocol
    ...sequencing primers, including: Is the primer unique? Make sure the primer only anneals once within the entire...incorrect sequence. Look at the peaks in the area and make sure they are justifiable peaks. For instance, ...

  47. Protocol - Bacterial Transformation

    Type
    Protocol
    ... that increase the transformation efficiency and make bacteria more susceptible to either chemical or ... plasmids at once. To save money, many labs also make their own competent cells. This is a relatively ...

  48. Lentivirus Production

    Type
    Protocol
    ...endonuclease negative E. coli strain such as NEB stable. Make a mixture of a total of 500 μL PEI-OptiPro SFM with...-Tip There can be batch to batch variation when making the PEI working stock, therefore the ratio of μg...

  49. Pipetting Protocol

    Type
    Protocol
    ...plunger. Remove the entire pipette from the container making sure not to touch the sides of the container with...important for scientists to calibrate their pipettes to make sure that they are properly tuned to dispense the...
Showing: 51 - 100 of 802 results