...remove the plate seal from the 96-well plate and set aside. Aspirate the wells and use a multichannel ...the plate. Place the plate on a microplate shaker set at 400 rpm and shake for 1 min . Repeat steps 8–10...a plate seal and incubate on a microplate shaker set to 400 rpm for 2 h at room temperature or overnight...the plate Place the plate on a microplate shaker set at 400 rpm and shake for 1 min . Repeat steps 18–...

...water bath on and set the appropriate temperature as described in your protocol. How to set the temperature...dial or a digital display you can set to the correct temperature. Set up your water bath so that it will...also helps the water bath heat up quicker. Pro-Tip Set up the water bath 30 minutes to 1 hour before you...

...Without moving past the maximum possible setting on the pipette, set the desired volume of the liquid on the...tip is properly set on the pipette to ensure pipetting accuracy. Once the pipette is set and you have the... pipette tip, the volume display which shows the set volume for the the pipette, and the tip ejector ring...

...PCR machine. Set annealing temperature 5°C below the primer melting temperature (Tm). Set extension step...create thousands of identical copies using a simple set of reagents and a basic heating and cooling (denaturing...choosing primers. Place thin-walled PCR tubes on ice Set up a 50 μL reaction (Keep all your reagents on ice...designed to have similar melting temperatures (Tm). Set the annealing temperature to 5°C lower than the Tm...MgCl 2 and/or 1µl DMSO to each reaction. The ideal setup for this troubleshooting step is to do one reaction...

...multichannel pipette set to 5 µL. For dilutions 6–8, use the 20–200 µL multichannel pipette set to 50 µL. For...dilutions, use the 20–200 µL multichannel pipette set to 90 µL and mix by pipetting the liquid up and down...ddPCR plate onto a chilled 96-well freezer block and set aside in the droplet generation BSC to cool. Prepare... Droplet Reader. Open the QuantaSoft software to set up a new plate layout. Designate the sample name,...

...the recipient plasmid, it is useful to see if one set of enzymes will work in the same restriction enzyme...future steps. Experimental Procedure Digest your DNA Set up restriction digests for your donor and recipient...recipient plasmid to insert. It is also important to set up negative controls in parallel. For instance, a...

...tubes in place, and the control panel where you can set the time and speed needed for your experiment. Procedure... close the external lid. Adjust the centrifuge settings according to speed and time (and temperature, ...

...: Standard Insert + Vector DNA Ligation Before setting up the ligation reaction itself, it is important... 20μL for storage at -20°C. Whenever you need to set up ligations in the future you can thaw a new tube...

... no additional water (<5 μl). It is advisable to set up multiple reactions by holding the vector concentration...insert, thereby increasing your chances of success. Set up a vector only control with water instead of the...

...Estimate of time required: ~3 h Workflow Timeline Plate set-up: 2 h qPCR run: 1.5 h Data analysis: 30 min Equipment...load the standards and samples onto the qPCR plate Set up and load the 96-well plate: Load 5 μL of each ...repeat 39x from step 3 / melt curve Example of plate set-up: 1 2 3 4 5 6 7 8 A 1.00 x 10 9 1.00 x 10 8 1.00...

...spectrophotometer 37 °C, 5% CO 2 incubator with shaking platform set to 120 rpm 37 °C bead bath Clamp stand and clamps... NanoDrop Spectrophotometer using the A280 IgG setting. Combine all eluates with measurable protein. Determine... NanoDrop Spectrophotometer using the A280 IgG setting. If the concentration of the pooled sample is above...

...remove virus from pipette tip. To mix the dilution, set the P200 pipette to 200 µL and pipette 10–20 times...ddPCR plate onto a chilled 96-well freezer block and set aside in the BSC to cool. Prepare the RRE/RPP30 master... Droplet Reader. Open the QuantaSoft software to set up a new plate layout. Designate the sample name,...calculate the titer. Calculations In the experimental setup above, the following virus dilutions were used to...factor v = Virus volume, mL *In the experimental setup above, the cells per well is 300,000 and the virus...

...counter 37 °C, 5% CO 2 incubator with shaking platform set to 120 rpm 37 °C bead bath Vortex Stir bar Magnetic...a 37 °C, 5% CO 2 incubator on a shaking platform set to 120 rpm. Pro-Tip Do not use cells that are over...into a clean microcentrifuge tube. Pro-Tip Cells settle quickly and need to be resuspended before sampling...

...ready for restriction digestion. Digest Your DNA Set up restriction digests for your PCR product and recipient...recipient plasmid to insert. It is also important to set up negative controls in parallel. For instance, a...

...and your initials. Labeling within a laboratory setting is important for organization, and it is recommended...

...measurement. Hit the tare button (or zero button) to set the balance to zero. Pro-Tip If you’re weighing a...

...MfeI. With oligo overlap cloning, you can design a set of oligos containing your desired restriction sites...

...technique, including personal protective equipment, setting up a clean workspace, and maintaining sterility...

...microscope stage) Reagents None needed Procedure Set up your microscope by removing any protective cover...

...solidified. Pro-Tip If you are in a hurry, the gel will set more quickly if you place the gel tray at 4 °C earlier...increases the density of your DNA sample causing it settle to the bottom of the gel well, instead of diffusing...