Skip to main content
Addgene

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

In Category

We narrowed to 987 results for: igh

Showing: 851 - 900 of 987 results

  1. Protocol - How to Run an Agarose Gel

    Type
    Protocol
    ...Casting tray Well combs Voltage source Gel box UV light source Microwave Reagents TAE ( recipe here ) Agarose...you to visualize the DNA under ultraviolet (UV) light. Caution EtBr is a known mutagen. Wear a lab coat...gauge how far the DNA has migrated; 2) it contains a high percentage of glycerol that increases the density... in the first place. Carefully load a molecular weight ladder into the first lane of the gel. Note: When...the second stop and carefully raise the pipette straight out of the buffer. Carefully load your samples...destain for 5 mins. Using any device that has UV light, visualize your DNA fragments. The fragments of ...minimize damage to the DNA. Note: When using UV light, protect your skin by wearing safety goggles or ...

  2. Plasmid Cloning by PCR (with Protocols)

    Type
    Protocol
    ...are having trouble getting colonies, you might want to use higher competency cells. Additionally, if your...amplify your insert DNA. It is important to use a high fidelity taq polymerase to minimize mutations. The...the digest goes at least 4 hours and as long as overnight. If you are going to use only one restriction ...final product is going to be very large (>10kb) you might want to use electro-competent cells instead of the...insert plate that are not correct. If you have a high number of colonies on your recipient plasmid alone...more colonies you will need to pick) and grow overnight cultures for DNA purification. After purifying... by Sequencing PCR based cloning carries a much higher risk for mutation than restriction enzyme based...

  3. General Transfection

    Type
    Protocol
    ...controller Hazardous waste container Reagents DMEM high glucose, Corning 10-013-CV L-alanyl-L-glutamine ...alternative such as glutaGRO) To a 500 mL bottle of DMEM high glucose, add 55 mL of heat-inactivated FBS and 5...health of the cell line is critical for obtaining high levels of virus. HEK293T cells should be split 3...purification should include an endotoxin removal step. For high quality plasmid DNA, the plasmid should also be ...transfection to determine what ratio gives the highest percentage of GFP positive cells. Refer to the ...pRosetta :µg of PEI. The 1:2 and 1:3 ratios provided high transfection efficiencies as can be seen here by...green fluorescent protein expression (green in the right panels) with a limited effect on cell growth. Last...

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

    Type
    Protocol
    ...are having trouble getting colonies, you might want to use higher competency cells. Additionally, if your... on your gene of interest (YGOI for short). You might need to express YGOI in cultured mammalian cells...the digest go at least 4 hours and as long as overnight. If you are going to use only one restriction ...final product is going to be very large (>10kb) you might want to use electro-competent cells instead of the...insert plate that are not correct. If you have a high number of colonies on your recipient plasmid alone...more colonies you will need to pick) and grow overnight cultures for DNA purification. After purifying...

  5. Centrifugation

    Type
    Protocol
    ...significantly higher density than water, then you may need to fill the blank based on weight instead of ..., which is generated by spinning the sample at a high speed. Being able to separate solids from a liquid...Centrifuges sit on the floor and can be about bench height. They often spin at similar speeds to their smaller...larger containers, but they can also rotate at much higher speeds and are used for more specialized techniques...distributed evenly around the center (see image to right). Even distribution ensures that the centrifuge ...

  6. Protocol - Bacterial Transformation

    Type
    Protocol
    ...will often get higher transformation efficiencies with less DNA, especially when using highly competent cells...transformation efficiencies upon thawing. For the highest transformation efficiency, we recommend that you... for the purposes of storage and amplification. Higher efficiency cells are more important if you will...won't grow in colonies. Incubate plates at 37°C overnight. Tips and FAQ How can I save time when carrying...reduce the efficiency of the transformation, so when higher efficiency is needed follow the complete protocol...

  7. AAV Titration by qPCR Using SYBR Green Technology

    Type
    Protocol
    ...Use a "Universal" SYBR master mix which contains a high-quality DNA polymerase and a blend of dTTP/dUTP ... use plasmid #59462 from Addgene. The values highlighted below in red were calculated using this plasmid...of Addgene plasmid #59462 : 1.07 μg/μL Molecular Weight: 6208 bp x 650 dalton/bp or (g/mole)/bp = 4.03 ...Dilution 8 20 uL Dil. 7 80 uL 5X 3125000X Dilutions highlighted in green are the ones loaded onto the qPCR plate...

  8. Western Blot

    Type
    Protocol
    ...acrylamide when immunoblotting high molecular weight proteins and higher percentage acrylamide when immunoblotting...protocol may need to be adjusted to accommodate slight differences between products. Addgene does not ...immunoblotting low molecular weight proteins. This protocol uses a dry transfer device but can be adapted...typically between 1–10 μg/mL. Incubate the membrane overnight in primary antibody at 4 °C on a rocking platform...

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

    Type
    Protocol
    ...subtract the weight of the empty tube from the weight of the tube with the gel fragment. The weight of the ...gel in a labeled microfuge tube. Using a scale, weigh the tube with the gel fragment after zeroing the... the gel percentage to get better separation. A higher percentage agarose gel will help resolve smaller...

  14. 'Tis the Season to #DeckTheLab

    Type
    Blog Post
    ... annual #DeckTheLab contest. The bar is set very high from last year’s impressive entries, but we have...

  15. The Twelve Days of Antibodies

    Type
    Blog Post
    ... to me… Antibodies developed by IPI!   🎶On the eighth day of Antibody, Addgene gave to me…  A recombinant...

  16. Protocols for Molecular Biology, Plasmid Cloning, and Viral Preps

    Type
    Protocol
    ...sample Using a Light Microscope Learn about the parts of a light microscope and its use Weighing Reagents Learn...applications, with videos for select protocols in the right-hand column. You can find even more video content... Learn how to weigh laboratory materials on a balance Basic Molecular Biology Name Description (Link opens...

  18. The 12 Days of CRISPR: 2021

    Type
    Blog Post
    ...editing by a miniature CRISPR-Cas12f nuclease. On the eighth day of CRISPR, Addgene gave to me: the PEDAR strategy...

  20. #Deckthelab Contest Winners 2017

    Type
    Blog Post
    ...annual #Deckthelab contest. The bar was set very high from last year’s impressive entries, but our community...

  22. Twenty Years of Sharing Science!

    Type
    Blog Post
    ...in 8,900 institutions across 111 countries. Just eight days into 2024, we've already shipped 2,078 items...

  24. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    ...controller Hazardous waste container Reagents DMEM high glucose, Corning 10-013-CV L-alanyl-L-glutamine ...alternative, such as glutaGRO) To a 500 mL bottle of DMEM high glucose, add 55 mL of heat-inactivated FBS and 5mL...sample of possible dilutions. You may want to try higher/lower dilutions depending on your downstream applications...transduced with lower dilutions of the virus will have higher levels of expression. Consider expanding populations...population may drop. This is because cells that express high levels of the transgene may have reduced growth ...

  31. Isolating a Monoclonal Cell Population by Limiting Dilution

    Type
    Protocol
    ...counter Reagents Polyclonal stable cell pool DMEM high glucose, Corning 10-013-CV L-alanyl-L-glutamine ...alternative, such as glutaGRO) To a 500 mL bottle of DMEM high glucose, add 55 mL of heat-inactivated FBS and 5...for one 96-well plate. Allow the cells to grow overnight. Replace the medium on the cells and collect the...conditioned medium, it will be more accurate if a higher volume of homogenized cell solution is transferred... Western blotting to screen for lines with the highest or lowest transgene expression ( Figure 2 ). Sample...

  36. Summer Fun at Addgene!

    Type
    Blog Post
    ...finally warming up in Boston and that means summer is right around the corner. Here at Addgene, we’re always...

  37. Our New Antibody Mascot is....

    Type
    Blog Post
    ...winner will be determined by popular vote. That’s right - you get to choose the new name! Additionally, ...

  40. New Viral Vectors - Spring 2025

    Type
    Blog Post
    ...interneurons. Take a look to see if these new preps might enhance your research!   Table 1: New ready-to-use...

  44. Editor's Choice, October 2016

    Type
    Blog Post
    ...for detecting genome edits, to discover why you might want to use Reddit to discuss your science and more...

  46. AAV Production in HEK293 Cells

    Type
    Protocol
    ...PES. Using a PES filter will maximize titer . DMEM high glucose, Corning 10-013-CV DMEM, low glucose (1 ...alternative, such as glutaGRO) To a 500 mL bottle of DMEM high glucose, add 55 mL of heat-inactivated FBS and 5...dissolved. Pro-Tip This step is challenging due to the high viscosity of PEG. Stirring under medium heat will...precipitation. Precipitation of the viruses can proceed overnight at 4 °C if needed. Transfer the entire sample ... prep . The clarified supernatant can be kept overnight at 4 °C before proceeding with the purification...

  47. Protocol - How to Create a Bacterial Glycerol Stock

    Type
    Protocol
    ...Inoculating an Overnight Liquid Culture . After you have bacterial growth, add 500 μL of the overnight culture...bacteria onto an LB agar plate . Grow your bacteria overnight at the appropriate temperature. Growth conditions...page. The next day you will be able to start an overnight culture for plasmid DNA prep the following day...
Showing: 851 - 900 of 987 results