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Addgene

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

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We narrowed to 602 results for: sam

Showing: 541 - 560 of 602 results

  10. Chemogenetics Guide

    Type
    Guide
    ...arePharmacologically Selective Actuator Modules (PSAMs, pronounced SAMs). PSAMs are engineered α7 nicotinic acetylcholine...varenicline. This PSAM carries mutations at L131G, Q139L, and Y217F and is termed PSAM4 (or PSAM 4 ). Additional...cognate PSEM. For example, PSAM Q79G,Q139G is activated by PSEM 22S , while PSAM L141F,Y115F is activated... effort to develop a PSAM that responded to a clinically approved drug, the PSAM domain was engineered...have also been developed as agonists for PSAM4. A given PSAM is coupled with an ion pore domain (IPD) ...signaling. PSAM-based LGICs can be classified by the ion pore domain they are coupled to: PSAM-Gly LGICs...ions, and inhibits neuronal activity. PSAM-5HT3 LGICs pair a PSAM domain with a 5-HT 3 serotonin receptor...

  11. 27 Hot Plasmids from 2016

    Type
    Blog Post
    ...population within a given tissue rather than a mixed sample. TRIBE was developed in Drosophila, and further...

  12. AAV Titration by qPCR Using SYBR Green Technology

    Type
    Protocol
    ...empty NTC C AAV reference Sample 3 D E Sample 1 Sample 4 F E Sample 2 Sample 5 F Perform data analysis...most samples. If sample is expected to have a titer <1 x 10 12 GC/mL, use dilutions 3–6 If sample is expected...standards and samples in duplicate at least If possible, include an AAV reference sample of known titer...Count the number of samples (n) and prepare master mix for an additional 10 samples (n+10 – the additional...that there is enough master mix for all samples). Each sample requires 15 μL of master mix. Pro-Tips Use...pipetting error and sample evaporation. Make the master mix after all the samples have been added to the...virion). 5 μL sample + 39 μL H 2 O + 5 μL 10X DNase buffer + 1 μL DNase Gently mix sample (do not vortex...

  13. Centrifugation

    Type
    Protocol
    ...tube filled with water to the same volume as your samples. If your samples are significantly higher density...liquid sample. It achieves this by using centrifugal force, which is generated by spinning the sample at ... which you need to spin your samples. For example, spinning down samples in 1.5 mL microfuge tubes at ...the pellets form in approximately the same place in each sample. Place the centrifuge’s internal lid on...centrifuge to separate different components in a liquid sample. Protocols... containers, spin at different speeds, or keep samples at specific temperatures. This protocol will cover...for the biosafety level of the lab space Reagents Sample to be centrifuged Water Background Information ...

  14. Using a Light Microscope Protocol

    Type
    Protocol
    ...helps concentrate the light onto the sample, and then through the sample. However, some of that light won’...hitting the sample or b) using dyes or stains that add color to some components of the sample but not others... power objective to find your sample. Place your slide (or other sample type) on the microscope stage....so that the light shines onto your sample. To ensure that your sample is adequately illuminated, look through... coarse focus knob to bring your sample into focus. Once your sample is in focus, use the stage arm to... you can easily see your sample. Then use the focus knobs to bring the sample into focus. Pro-Tip As you...magnification used, any stains used in your sample, and sample information so that you or others can interpret...

  15. Coomassie Purity Stain of Recombinant Antibodies

    Type
    Protocol
    ...of 4X sample buffer to each sample. Add 2 µL 10X reducing agent to each sample. Spin the sample briefly...concentration of your samples based on your sample’s HC + LC area. Example calculation: If the sample HC + LC Area...Invitrogen NP0001 4X NuPage sample buffer, Invitrogen NP0007 10X NuPage sample reducing agent (500 mM dithiothreitol...microcentrifuge. Heat the samples for 10 min at 100 °C in a heat block. Spin the sample briefly in the microcentrifuge...one lane before loading samples. Load 20 µL of each recombinant antibody sample to the appropriate well... mobility of the bands (sample AR0016 in Figure 1) indicates that the samples may not have been processed... SimplyBlue SafeStain) and ImageJ software. The sample is separated by denaturing polyacrylamide gel electrophoresis...

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

    Type
    Protocol
    ...finite binding capacity. If your sample exceeds the capacity, divide the sample among multiple columns. For...4 mL of recombinant antibody sample. Pro-Tip If the volume of the sample is greater than 4 mL , then divide... Slowly apply the sample to the center of the compact resin bed. Pro-Tip For sample volumes <1.5 mL top...the sample with 0.2 mL PBS to aid protein recovery. Allow the resin bed to fully absorb the sample. Centrifuge...Spectrophotometer to see if the sample has reached the desired concentration. If the sample concentration is still... to see if the sample has reached the desired concentration. Gently transfer the sample from the reservoir...Determine the protein concentration of the pooled sample on the NanoDrop Spectrophotometer using the A280...

  17. Kit Free RNA Extraction

    Type
    Protocol
    ... 10 seconds. Incubate sample(s) for 15 minutes on ice and centrifuge the sample(s) for 15 minutes at 12,000...Always wear gloves, and whenever possible, keep RNA sample and reagents cold and work quickly to reduce RNA...use 1 mL of Solution D per 1 X 10 7 cells. Allow sample(s) to sit at room temperature for 5 minutes to ... tissues such as bone, or bacteria/yeast/plant samples will require additional steps to effectively lyse...open the cells. Extract RNA from the homogenized sample(s).Transfer tissue/cell lysate to a 4 mL tube. ...of the tissue/cell lysate. Pro-Tip Having your samples spin at 4°C helps reduce RNA degradation. If you.... You may have to collect twice or more from the same tube, but unlike using a p1000 tip it will give ...

  18. Lentivirus ddPCR Titration

    Type
    Protocol
    ...concentration of each sample on a spectrophotometer. Prepare 25 ng/µL stocks of each sample. Samples can be used...tube as shown below. For 8 samples prepare enough master mix for 9 samples. Component Volume 9X Volume...constant across samples. In the RRE example below, 2-fold serial dilutions of a sample were loaded in ...ddPCR Lentivirus sample data, RPP30 positive (green) and negative (black) droplets Sample ID Plasmid ID ... higher dilutions depending on their particular sample. This protocol was modified from the publication...Starting Thaw the master mix, primers/probe mixes and samples on ice before use. Wipe down all pipettes and surfaces...C until ready to use. Preparing for ddPCR Thaw samples, primers/probe mixes, and master mix on ice. Before...

  19. Western Blot

    Type
    Protocol
    ...duplicate samples on the plate. Subtract the average absorbance of the blank from all of the samples. Plot...concentration of the sample from the standard curve. Determine the volume of sample required to load an...protein for each sample. Pro-Tip The ideal total protein loaded will vary between samples and target proteins...abundance in the sample you will need to load a greater amount of total protein. Prepare sample for loading...Dilute 4X protein loading buffer in the sample to 1X . Boil the samples for 10 min at 100 °C . Section 3: SDS-PAGE...blotting to visualize a protein from cells or tissue samples. Equipment Microcentrifuge 0.5–10 µL single channel...duplicate, dilute 10 µL of standard, blank, and lysate samples into 200 µL of BCA reagent in a 96-well microtiter...

  20. AAV ddPCR Titration

    Type
    Protocol
    ... ddPCR sample data Sample Dilution Concentration Titer (GC/mL) Average Titer (GC/mL) AAV Sample 1 6400000...tube as shown below. For 8 samples prepare enough master mix for 9 samples. ITR Master Mix Volume 9X Volume...prepared for each AAV sample, and the three final dilutions are assayed. The samples that are assayed are... higher dilutions depending on their particular sample. For additional tips on AAV titering using ddPCR...channel 1–10 µL pipette to add 5 µL of each viral sample to Dilution 1 in the 48-well dilution plate and...not push the cap in all the way, just ensure the samples are covered. Bring the PCR tubes to the BSC used...software to set up a new plate layout. Designate the sample name, experiment type, supermix type (ddPCR Supermix...
Showing: 541 - 560 of 602 results