We narrowed to 988 results for: IGH@

...PEI μg ratios provided high transfection efficiencies as measured by the highest proportion of GFP positive...without limiting cell growth. Left panels: bright field images; right panels: GFP channel images. Last reviewed...meter Stir plate Magnetic Stir Bar 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 ...the packaging cell line is critical for obtaining high viral titer. 293T cells should be split 3 times ...purification should include an endotoxin removal step. For high quality plasmid DNA, the plasmids should also be...transfection to determine what ratio gives the highest percentage of GFP positive cells. Refer to the ...

...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 ...

...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...

...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...

...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...

...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 ...

...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...

...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...

...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...

...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...

...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 These protocols are the ...

...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 ...

... homepage! You can track the number of plasmids right below the search bar and more stats further down...

...a: That will warm up the entire lab (hint: you might need some mini marshmallows…). We’ll choose 4 submissions...

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

...! Our mascot who has supported us over the past eight years now has a new friend to join them. Blugene...

...starting their careers in the lab and even those who might just need a refresher on some best practices for...

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

...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...

...Segment on mCyRFP1 Hot Plasmid 4: mScarlet - a new, bright, and monmeric red fluorescent protein.  Listen ...

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

...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...

...Goedhart  Created in collaboration with Addgene writers High Complexity Golden Gate Assembly with NEB's DADExperts...

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

...with you.  Top 5 science comics:   1)   “Piled Higher and Deeper” by Jorge Cham   2)   “Boxplot” by Maki...

...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...

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

...journey. And if you have additional resources that might be helpful, please reach out to me by email.  About...

...organized by graduate student researchers. That’s right, #ActualLivingScientists, eager to communicate their...

...topic page Read Joachim's Series on Choosing the B(right)est Fluorescent Protein Plasmids 101: Visualizing...

...plasmids at Addgene, along with other tools like high resolution Ras sensors and crosslinked peptides ...

...and career goals, or to answer any questions you might have about Addgene. The page is divided into three...

...the Expression and Insert Species filters on the right-hand side of the search page to narrow your keyword...

... University with the backing of a prestigious Fulbright Fellowship. Dr. Kwapata now works as a lecturer...

...container Inverted fluorescence microscope 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...each well of the 6-well dish. Incubate the cells overnight. If using freshly collected virus, filter through...If you anticipate that your viral stock will be higher titer, consider additional dilutions. Dilution ...

...shape Addgene's future. Listen to this episode for insights into the process of getting an MD PhD, the inside...

...back on some fascinating Addgene history that you might not have known about. Here's the schedule for the...

...plasmids at specific molar ratios. To get these ratios right, you need to do a few calculations. This calculator...

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

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

...of the contest is our traditional approach. You might decorate your lab in pipette tip snowflakes, hang...

...fluorescent protein family that give off color in ambient light. Expressing these chromoproteins in E. coli would...

... about Crystal’s exciting career path, to gain insight into industry research, and to get advice on how...

...protocol may need to be adjusted to accommodate slight differences between products. Addgene does not ...mg/mL DAPI solution to 100 µL PBS. Protect from light. Prepare a 300 nM DAPI working solution by diluting...DAPI stock solution into 5 mL PBS. Protect from light. Procedure Section 1: Seeding cells Place a sterile...Pro-Tip The plate can be wrapped in foil to block light. Remove the secondary antibody and dispose of it...

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

...open to industry scientists. This collection is highly diverse in terms of target organisms (from bacteria...

... school for microbiology, and I ended up going straight to law school. This was in the early 80s, and ...

...plasmids shipped. Reaching one million plasmids shared might have you wondering how many plasmids leave Addgene...

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

...the labs they’re granted to, as well as shine a light on some of the great minds working in developing...