...generally considered biosafety level 1 but may require BSL-2 handling depending on the insert. Please ...liquid up and down 10–20 times. Gently cover the entire dilution plate with Microseal adhesive seal - do...droplets are not disrupted, insert the pipette tips directly in the center of the well and tilt to a 45° angle...

...bacterial colonies. Equipment Sterile toothpicks or wire loop Bunsen burner (or other small flame source)... top of the glycerol stock. Pro-Tip If you use a wire loop you can sterilize it by passing it through ...

...and with a clean, sterile razor blade, slice the desired DNA fragment from the gel. Notes: To protect the...with the gel fragment. The weight of the gel is directly proportional to its liquid volume and this is ...

...you can design a set of oligos containing your desired restriction sites and add them to your existing...overlapping oligos that once annealed can be cloned directly into the overhangs generated by restriction digest...

...Set up your water bath so that it will reach the desired temperature by the time you need it for your experiment...water bath to prevent evaporation and maintain the desired temperature. This also helps the water bath heat...

...cells Antibody Validation Using the Indirect ELISA Method Run an indirect ELISA against a purified antigen...

... likely to ligate to itself rather than to the desired insert. If you are in this situation, it is important...Additional controls are encouraged, but may only be required for troubleshooting failed ligations. The following...

...maximum possible setting on the pipette, set the desired volume of the liquid on the pipette. Depending ... you can slowly release the plunger. Remove the entire pipette from the container making sure not to touch...

...to the template DNA and the Taq polymerase, PCR requires free nucleotides [dNTPs; adenine (A), cytosine... thymine (T)] in an equal molar ratio. It also requires two unique single stranded DNA oligonucleotide...

...Last Update: February 13, 2019 Estimate of time required: ~3 h Workflow Timeline Plate set-up: 2 h qPCR...enough master mix for all samples). Each sample requires 15 μL of master mix. Pro-Tips Use a "Universal...

...recheck the pH to prevent over or undershooting the desired pH. Allow the solution to mix for 10 min and then...proceed overnight at 4 °C if needed. Transfer the entire sample to 3 x 500 mL conical bottles and centrifuge...

...plasmid DNA, the plasmid will already be in your desired bacterial strain and you will not need to obtain...

...wear glasses/goggles in addition to the BSL-1 requirements. Conclusion Although simple, following appropriate...

... in LB medium. The concentration of antibiotic required for effective over-agar selection has been empirically...

...balance and then tare it. Then add the material directly to the tube to weigh it. Make sure that your scoopula...

...linearize a plasmid to determine the size of the entire construct or excise some or all of an insert from...

...The annealed but nicked vector product is then repaired during the replication cycle. Empty vectors for...

...the cells in the remaining well, a cell count is required for calculating the titer. Incubate for 48–72 ...

... ∼500 ng of DNA, while molecular cloning often requires 1 µg of DNA. The total reaction volume usually...

...recheck the pH to prevent over or undershooting the desired pH. Allow the solution to mix for 10 min and then...