...livestock diseases such as malaria and toxoplasmosis. Despite their importance to global health, most apicomplexan...

... Harvest Gently aspirate media from the 6-well plates. Be sure to use a new aspirating pipette for each... 37 °C for 30 min. Gently aspirate media from wells, using a new aspirating pipette for each well. Wash...Wash wells with 1 mL of 1X PBS and aspirate, using a new aspirating pipette for each well. Detach cells...Equipment Class II, Type A2 Biological Safety Cabinet Aspirating unit Microcentrifuge, Eppendorf, 5425R Droplet...tube. Centrifuge for 5 min at 100 x g . Gently aspirate supernatant. Wash cell pellets in 500 µL PBS. .... Centrifuge for 5 min at 100 x g . Gently aspirate supernatant. Extract genomic DNA according to the ...primers/probe mixes and master mix for 15 sec then spin 10 sec in a mini centrifuge and place on ice. Wipe...

...plate seal from the 96-well plate and set aside. Aspirate the wells and use a multichannel pipette to add...a total of three washes. After the final wash, aspirate the wash buffer from the wells. Prepare the blocking...a total of three washes. After the final wash, aspirate the wash buffer from the wells. Prepare antibody...a total of three washes. After the final wash, aspirate the wash buffer from the wells. Dilute the HRP-conjugated...

...the speed at which you need to spin your samples. For example, spinning down samples in 1.5 mL microfuge...using centrifugal force, which is generated by spinning the sample at a high speed. Being able to separate...that can accommodate different sized containers, spin at different speeds, or keep samples at specific... floor and can be about bench height. They often spin at similar speeds to their smaller cousins, but ... if applicable) listed in your protocol. Pro-Tip Spin speed is often given as Relative Centrifugal Force...and investigate the issue. When you are finished spinning your samples, carefully remove them from the centrifuge...

...NaCl PBS. Spin at 3000 rpm for 5 min at 4 °C and discard the flow through. Add your sample. Spin at 3500...0.1% Pluronic F68 PBS and incubate for 10 min at RT Spin at 3000 rpm for 5 min at 4 °C and discard the flow... Pluronic F68 PBS and incubate for 10 min at RT. Spin at 3000 rpm for 5 min at 4 °C and discard the flow...C, discard the flow through. Add more sample and spin 3500 rpm for 2–5 min at 4 °C, discard the flow ...that iodixanol is not easily removed. After each spin, add more formulation buffer and sample and make...

... NNGRR(N) Acidaminococcus sp. (AsCpf1) and Lachnospiraceae bacterium (LbCpf1) 5' TTTV AsCpf1 RR variant...

...pipette 2–20 µL single channel pipette Reagents Aspirating pipette, VWR 414004-265 5 mL pipette, VWR 89130...Sigma L2884 Aprotinin, Millipore Sigma A6279 Zeba Spin Desalting Columns, 7 kDa MWCO, 10 mL, Thermo Fisher... to Option 1 with a buffer exchange using a Zeba Spin 7 kDa MWCO desalting column. If the concentration...exchange using a desalting column The 10 mL Zeba Spin Desalting Columns can accommodate up to 4 mL of ...divide it into 2 columns. Twist off the 10 mL Zeba Spin Desalting Column’s bottom closure and loosen cap...

...protect individuals from potential accidents such as spills. BSL-2 is different because it includes labs that...to protect your body and clothing from potential spills. Gloves should fit well enough to be snug to your...your sleeves into your gloves to prevent potential spills down your sleeve. This may or may not be possible...worn to protect your legs and feet from potential spills. If wearing open-toed shoes, cloth coverings or...

... wells, aspirate the media from the wells. Gently wash the cells with 1 mL of PBS and aspirate wash. Filter...added until 48–72 h after transduction. Gently aspirate the media from the cells. Add 1.5 mL of DMEM complete...colonies should be visible. Every 3–4 days, gently aspirate the media and replace it with fresh DMEM complete...

...Vortex primers, probe and master mix for 15 sec then spin 10 sec in a mini centrifuge and place on ice. Wipe...Volume 20 µL Vortex the master mix for 15 sec and spin in a mini centrifuge for 10 sec before use. Place...generated, use a 20–200 µL multichannel pipet to aspirate 40 µL of droplets. Pro-Tip To ensure that the ...45° angle. Count to 20 while slowly and gently aspirating the droplets. Transfer the droplets to a prechilled...

...each T-175 flask: Aspirate culture media and rinse once with 10 mL of PBS. Aspirate PBS and add 2 mL of...at ~80% confluence. Aspirate culture media and rinse once with 60 mL PBS. Aspirate PBS and add 35 mL of...

...Untransduced 0 1498.5 1.5 Mix the dilutions well Gently aspirate media from the cells. Add 1.5 mL of a viral dilution...calculating the titer. Incubate for 48–72 h. Gently aspirate media and replace with 1 mL of PBS. Calculate ...

... hood. Clean any spills immediately, and decontaminate as necessary. For large spills that cannot be easily...

... the cells at 37 °C, 5% CO 2 for ~20 h. Gently aspirate media, add 10 mL fresh DMEM complete containing...following morning. The following morning, carefully aspirate the media. Replace the media with 15 mL of DMEM...

...the media and chloroquine and mix well. Gently aspirate the media out of the previously seeded 10 cm plate...following morning. The following morning, carefully aspirate the media. Replace the media with 10 mL of DMEM...

...sample. Add 2 µL 10X reducing agent to each sample. Spin the sample briefly in the microcentrifuge. Heat ...the samples for 10 min at 100 °C in a heat block. Spin the sample briefly in the microcentrifuge. Remove...

... tissue/cell lysate. Pro-Tip Having your samples spin at 4°C helps reduce RNA degradation. If you don’...

...100 μg/mL ampicillin or carbenicillin. Day 2: 2. Spin down the cultures and use a miniprep kit to obtain...media from cells and pool with media from Day 4. Spin media at 1,250 rpm for 5 minutes to pellet any HEK...

...Repeat for each sample. Notes: Keep in mind that despite the accuracy of the NanoDrop, if two consecutive...

...transfer it into your container. Make sure that any spills produced during the protocol are cleaned up. Discard...

...Section 2: Fixing and permeabilizing cells Gently aspirate the media from the 24-well plate. Wash each well...

...isopropanol to precipitate the plasmid DNA. Either spin to pellet the DNA or apply the solution to a column...

...Incubate the cells with the virus for 48–72 h. Gently aspirate the media from the cells. Add 1.5 mL of DMEM complete...

... bit more than 200 mL (~220 mL) just in case we spill anything or have small errors in measurement. You...