We narrowed to 821 results for: EED

... Ligation Independent Cloning (LIC) obviates the need for the time-consuming ligation step of traditional...protocol ). After the digest is complete, you will need to separate the linearized vector from the reaction...free nucleotides from your PCR product before proceeding, as they will interfere with the exonuclease ...

... biosafety regulations. Workflow Timeline Day 0: Seed 293T cells Day 1: Transduce cells Day 2 (am): Remove...supernatants to multiple freeze-thaw cycles. Procedure Seed 75,000 cells into each well of a 6-well dish. Prepare...integration event per cell. When the percentage exceeds 40% you risk counting cells with multiple integration...

... this method extensively. Why Gibson Cloning? No need for specific restriction sites. Join almost any ... plasmid, think about what DNA segments you will need to join to create your final plasmid. Adjacent segments...Stitching” fragments together using oligos When you need intervening sequence between two PCR products, one...

...broken glass or needles. Sharps containers must be thick walled, impenetrable by a needle, and must be able...cholerae . BSL-2 includes all of the precautions needed in BSL-1, along with additional precautions to ...

...5, 10 or 20μL for storage at -20°C. Whenever you need to set up ligations in the future you can thaw a...ligation can be improved by incubation at 37°C. Proceed with bacterial transformation . Tips and FAQ Do...of DNA no matter how long it is, and therefore we need to adjust the amount of DNA used in a ligation based...

...that can fit on a microscope stage) Reagents None needed Procedure Set up your microscope by removing any...of view is centered on a region of interest. If needed, you can also use the fine focus knob (the smaller...objective! Once on the higher objective, you will likely need to re-adjust lighting and focus. Increase the power...

...Centrifuge the culture to pellet the bacteria before proceeding with DNA preparation. Pro-Tip If your entire ...DNA to precipitate. Centrifuge solution at high speed (at least 12,000 rpm) for 15-30 min at 4°C. Notes...from the DNA pellet. Centrifuge solution at high speed (at least 12,000 rpm) for 5 min at room temp. Pour...

... GGC TGT TAG AGA GAT AAT TGG A 3’). TIP: You may need to adjust the sequencing conditions if the DNA polymerase...scaled to produce different amounts of virus as needed. Day 1: a. For each plasmid to be transfected, .... Change to fresh puromycin-containing media as needed every few days. h. Assay infected cells. The following...plastic pipettes in place of glass pipettes or needles. Liquid waste should be decontaminated with at ...

...protein content of the sample is determined. Sharing speeds science. We believe that sharing the full details... However, please be aware that your protocol may need to be adjusted to accommodate slight differences...CSV file. Select import. Choose file. Format as needed. Select the Insert tab, then Chart . This will ...

...in your desired bacterial strain and you will not need to obtain more competent cells and retransform. ...want to store bacteria for a longer time, you will need to establish glycerol stocks. The addition of glycerol...

...differing in their resistance genes, as one does not need to prepare separate batches of antibiotic-containing...experiment. For publishable data, the experiment would need to be repeated to account for variability....

...concentration of the DNA that you isolated before proceeding to your next intended step with the now gel purified...want to load on a gel, make 10% more volume than needed because several microliters can be lost in pipetting...

...Summary Oligo overlap cloning can be used anytime you need to add a short stretch of DNA to a plasmid, such...GTATAC AATTAATT CCGCGCGG GTTAAC - 5' = 28 bp We also need to include additional bases to complement the overhangs...

...Before beginning your diagnostic digest, you will need to select a restriction enzyme or enzymes that cut... vector by a single restriction enzyme, you will need to verify that it has be cloned in the correct orientation...

... which minimizes the number of small volumes you need to pipet. If you are using the same template DNA...them as a lyophilized powder. The researcher then needs to reconstitute their primers in liquid, normally...

...gel-ager mix than you think you’ll need. So for 220 mL of LB-agar, we’ll need: (37 g pre-mixed LB-agar powder...

...the physical number of viral particles. Users may need to run lower or higher dilutions depending on their...Seal’ button. After the plate has been sealed, proceed to thermocycling. Thermal Cycling Run the following...

...concentration of a substance based on its absorbance, you need to know the wavelength of light that your substance...

...results? If you require additional sequencing and need to design a custom primer, Addgene recommends using...

...pipette tips can be sterilized in an autoclave as needed to prevent contamination. When not in use, the ...