...for the presence of particles still in solution. Continue to stir until all particles have dissolved. ... bottle and stir on a magnetic stir plate. Stir until all particles have dissolved. Note: The powder should...µm filter. Prepare aliquots and store at -20 °C until use. BCD TFX 1000 mL BalanCD HEK293 Media 10 mL ...Glutagro Do not add selective reagents Store at 4 °C until use. We suggest preparing fresh solutions after ...HEK293 Feed 20 mL 200 mM Glutagro Store at 4 °C until use. We suggest preparing fresh solutions after ...Purification Protocol ). Store the supernatant at 4 °C until ready to use....

...water. While stirring, slowly add hydrochloric acid until the solution clears. Check the pH of the solution...adjust to a final volume of 1000 mL. Stir at RT until fully dissolved. Pro-Tip This step is challenging...poured on them) and can be adjusted by carefully tilting the CS5 back and forth. Pro-Tip To help distribute...distribute the media amongst the five layers, tilt the CS5 such that the media goes toward the cap. If the ...Mix well between rounds of sonication. Sonicate until no live cells can be seen when stained with Trypan...

...electrocompetent cells on ice for 15-20 minutes or until completely thawed. Chill a box of 200 µL micropipette...tube). Distribute evenly with a sterile spreader until all liquid has been absorbed by the agar. This usually...drips onto the lid. Let plates remain agar side up until dried before overnight incubation if needed. Place...addition of 10 mL cold LB and scrape for each plate until agar is clear (all bacteria have been removed). ...recovery media!). Do not proceed with Maxipreps or NGS until adequate transformation efficiency is obtained. ...

.... You can take entire courses on microscopy and still have more to learn, so, for this protocol, we'll...adjustment knob (the larger of the two focus knobs) until you can see the edges of the aperture are in focus...field of view and then slowly open the diaphragm until the edges are just out of view. Pro-Tip Looking ...arm to move the stage (and ultimately your slide) until your field of view is centered on a region of interest...

...buffers and do not use water). Microwave for 1-3 min until the agarose is completely dissolved (but do not ... OR let sit at room temperature for 20-30 mins, until it has completely solidified. Pro-Tip If you are...electrophoresis unit). Fill gel box with 1xTAE (or TBE) until the gel is covered. Pro-Tip Remember, if you added...additional wells of the gel. Run the gel at 80-150 V until the dye line is approximately 75-80% of the way ...

... Work in a well-ventilated space and under a fume hood when working with the volatile reagents in the ...entire tube is dried but while the white pellet is still visible. Resuspend RNA pellet in RNase-free water...

...digested DNA on 0.8% low melting point agarose gel until you can distinctly see 2 bands, one 7kb and one ... replace with fresh media. Do not add puromycin until at least 24 hours after infection to allow for sufficient...guidelines for the number of days you should wait until harvesting your cells. However, you should optimize...solution by dissolving 40g of LB powder in 1L of distilled water. Autoclave and cool to 55°C. Add 1mL of ...of puromycin (Sigma-Aldrich catalog #P8833) in distilled water. Sterilize by passing through a 0.22 μm ...

...Equipment Water Bath 10% bleach or 70% ethanol Distilled water Disinfectant Thermometer Water bath weights...the edge of the slide. Fill the water bath with distilled water, not tap water, to prevent salts from accumulating...water in it from previous uses. However, you will still need to regularly clean out the water bath even ...

.... Use the lysate immediately or store at -80 °C until ready to use. Section 2: Determine the total protein...power supply. Run the gel at 100 V for 10–15 min or until the samples have moved out of the wells and into...the voltage to 150 V and continue running the gel until you have obtained the desired separation. Gently...

...on the chilled 96-well freezer block in the BSC until ready to use. Prepare the Master Mix Place a ddPCR...pipette tips directly in the center of the well and tilt to a 45° angle. Count to 20 while slowly and gently...Lift the tips ~1 mm. Touch the side of the well and tilt the pipette tips at a 45° angle. Count to 20 while...

... used for ddPCR immediately or stored at -20 °C until ready to use. Preparing for ddPCR Thaw samples, ...pipette tips directly in the center of the well and tilt to a 45° angle. Count to 20 while slowly and gently...Lift the tips ~1mm. Touch the side of the well and tilt the pipette tips at a 45° angle. Count to 20 while...

... of the agar and gently spread over the surface until the liquid is mostly absorbed (there is a very small...onto the agar and gently spread over the surface until the liquid is mostly absorbed. The spreading of ...

...will remove bases from the 3' end of the cut site until the first G is reached (shown in blue), at which...causing the enzyme to perform exonuclease activity until the first "G" in the sequence. Because the polymerase...

...water. While stirring, slowly add hydrochloric acid until the solution clears. Check the pH of the solution...dislodge the cells. Incubate the cells for 18 h, or until the following morning. The following morning, carefully...

...water. While stirring, slowly add hydrochloric acid until the solution clears. Check the pH of the solution...disturb the cells. Incubate the cells for 18 h, or until the following morning. The following morning, carefully...

...large container holding a small volume of liquid, tilt the container holding the liquid so that it's easier...second stop to release the liquid. Pro-Tip Dispense until the first stop, wait, and then dispense to the second...

... with the recipient plasmid's MCS. However, you still need to avoid restriction enzymes that cut within...colonies are a result of uncut empty plasmid, you will still have colonies when you do not add ligase. If the...

...Transformation Summary PCR based cloning is incredibly versatile and allows for nearly any piece of DNA to be placed...colonies are a result of uncut empty plasmid, you will still have colonies when you do not add ligase. If the...

...the sample concentration is still too low, repeat the centrifugation until the volume of the sample is...

...been placed in the centrifuge before you and may still be on the equipment. Before using the centrifuge...