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Addgene

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

In Category

We narrowed to 304 results for: pET

Showing: 281 - 300 of 304 results

  1. Colony Formation Titering Assay

    Type
    Protocol
    ...single channel pipette 2–20 µL single channel pipette 20–200 µL single channel pipette 200–1000 µL single...single channel pipette Ice bucket CO 2 incubator Pipet controller Hazardous waste container Reagents DMEM...complete containing 10 μg/mL polybrene. Mix well by pipetting or inverting. Aliquot 1.35 mL of the cell suspension...

  2. Transfection for Recombinant Antibodies

    Type
    Protocol
    ...Biosafety Cabinet 4 °C Refrigerator Micropipettes and tips Pipettor and pipettes Benchtop centrifuge compatible...Reagent Preparation instructions. Wipe down all pipettes and equipment with 10% bleach prior to use. Reagent...Check the cell density and viability: Using a 5 mL pipette, transfer 0.5 mL of HEK293 cell suspension into...the microfuge tube containing the trypan blue. Pipette 10 times to mix. Load 10 µL of the cell suspension...

  3. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    ...single channel pipette 2–20 µL single channel pipette 20–200 µL single channel pipette 200–1000 µL single...single channel pipette Ice bucket CO 2 incubator Pipet controller Hazardous waste container Reagents DMEM...DMEM complete + 10 µg/mL polybrene. Mix well by pipetting or inverting the tube. Aliquot 1 mL of cell suspension...

  4. Isolating a Monoclonal Cell Population by Limiting Dilution

    Type
    Protocol
    ...single channel pipette 2–20 µL single channel pipette 20–200 µL single channel pipette 200–1000 µL single...single channel pipette CO 2 incubator Pipet controller Hazardous waste container Microscope Hemocytometer... by passing several times through a serological pipet or by passing through a 40 µm cell strainer mesh...

  5. Kit Free RNA Extraction

    Type
    Protocol
    ...storage in the cold room may damage it. Using a pipettor, transfer the top, aqueous phase to a new RNAse-free...disturb or collect the interphase layer with your pipette tip. Pro-Tip To collect as much of the top aqueous...interphase layer, consider using a lower volume pipettor like a p200 to collect a majority of the aqueous...

  6. General Transfection

    Type
    Protocol
    ...single channel pipette 2–20 µL single channel pipette 20–200 µL single channel pipette 200–1000 µL single...single channel pipette Ice bucket CO 2 incubator Pipet controller Hazardous waste container Reagents DMEM...

  7. Protocol - How to Run an Agarose Gel

    Type
    Protocol
    ...is unloaded, push the pipettor to the second stop and carefully raise the pipette straight out of the buffer...comb or towards the sides/edges of the gel with a pipette tip. Place newly poured gel at 4 °C for 10-15 mins... the tip. Place the very top of the tip of the pipette into the buffer just above the well. Very slowly...needed because several microliters can be lost in pipetting. For example, if you want to load 1.0 μg in 10...

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

    Type
    Protocol
    ...want to use electro-competent cells instead of the more common chemically-competent cells. The number of...Follow the manufacturer’s instructions for your competent cells. For most standard cloning, you can transform...transform 1-2μl of your ligation reaction into competent cells such as DH5alpha or TOP10. If using much...getting colonies, you might want to use higher competency cells. Additionally, if your final product is...

  9. Plasmid Cloning by PCR (with Protocols)

    Type
    Protocol
    ...want to use electro-competent cells instead of the more common chemically-competent cells. The number of...according to the manufacturer’s instructions for your competent cells. For most standard cloning, you can transform...transform 1-2μl of your ligation reaction into competent cells such as DH5alpha or TOP10. If using much ...getting colonies, you might want to use higher competency cells. Additionally, if your final product is...

  10. Video Library

    Type
    Protocol
    ...Mulitchannel Pipetting Technique Learn how to pipette multiple samples at once using a multichannel pipette and...and avoid inconsistent pipetting with our simple tips Bacterial Transformations We walk you through a standard...

  12. Molecular Biology Protocol - Restriction Digest of Plasmid DNA

    Type
    Protocol
    ...digest results. Equipment Electrophoresis chamber Pipetman Reagents Liquid DNA aliquot of your plasmid of...instructions) Gel loading dye Electrophoresis buffer Pipet tips Procedure Select restriction enzymes to digest....2-0.5 µL of enzyme because it is difficult to pipette less volume than this; 0.2-0.5 µL will likely be... more enzyme is usually better. Mix gently by pipetting. Incubate tube at appropriate temperature (usually...

  13. What is Polymerase Chain Reaction (PCR)

    Type
    Protocol
    ...minimizes the number of small volumes you need to pipet. If you are using the same template DNA for all ...wish to perform and add 10% extra to account for pipetting error. In this example, we have 7 different reactions... (0.14µl) = 1.54µl Mix the contents by gently pipetting up and down several times. Keep tube on ice. Add...mix so you know that the primers have been added, pipet the forward primer onto one side of the tube wall...

  14. Handling Plasmids from Addgene - Purifying Plasmid DNA

    Type
    Protocol
    ...chromosomes. Collect the supernatant into a new tube by pipetting or carefully pouring. Optional: Add 5 μL of 2 ...particles Bottom phase - Organic phase (protein) Pipet the aqueous DNA layer and place it in a new microfuge...the tube after the 70% ethanol wash. You can also pipet the supernatant out of the tube if you are worried...

  15. Pouring LB Agar Plates

    Type
    Protocol
    ... process works. Equipment Autoclave Water bath Pipetman Reagents 37 g pre-mixed powder consisting of: ...10 - 15 mL of agar. Autoclavable flasks Sterile pipettes Ice bucket to hold antibiotic Antibiotic at 1000...pouring. Measure your desired amount of agar with a pipete for the first plate to get a good idea of what ...

  16. Protocol - How to Create a Bacterial Glycerol Stock

    Type
    Protocol
    ...bacterial strain and you will not need to obtain more competent cells and retransform. Bacteria on an LB agar ... the tube and use a sterile loop, toothpick or pipette tip to scrape some of the frozen bacteria off of...

  17. Protocol - How to Streak a Plate

    Type
    Protocol
    ...stab or glycerol stock . Using a sterile loop, pipette tip or toothpick, touch the bacteria growing within...plate, to create streak #2. Using a third sterile pipette tip, toothpick, or sterilized loop, drag through...

  20. AAV Purification by Iodixanol Gradient Ultracentrifugation

    Type
    Protocol
    ... formulation buffer and sample and make sure to pipet back and forth a few times to mix the iodixanol ...buffer. Use a P1000 to the bottom of the filter and pipette up/down and wash off the walls of the filter to...
Showing: 281 - 300 of 304 results