...Making LB Agar Plates Create plates to culture bacteria in the lab Pouring LB Agar Plates Protocol Streaking...Bacteria on Plates Isolate single bacterial colonies on an agar plate Streaking Bacteria on Plates Protocol...Antibiotic Plating Create plates to culture bacteria in the lab Over-Agar Antibiotic Plating Protocol AAV...

... a shaking platform. Wash the membrane 3x for 5 min in 1X TBST at RT on a shaking platform. Section 6:... a shaking platform. Wash the membrane 3x for 5 min in 1X TBST at RT on a shaking platform. Prepare the...supply iBlot 2 Gel Transfer Device Roller Spatula Platform shaker Cold room Gel imager -80 °C freezer Reagents... Tween-20 Nonfat milk powder 96-well microtiter plate Chemiluminescence substrate Plastic wrap Primary...into 200 µL of BCA reagent in a 96-well microtiter plate. Incubate for 30 min at 37 °C . Determine the absorbance...average absorbance of the duplicate samples on the plate. Subtract the average absorbance of the blank from...overnight in primary antibody at 4 °C on a rocking platform. Pro-Tip Primary antibody incubation can be reduced...

...week: Monday: Plate 1×10 6 cells in a T75 flask in 15 mL DMEM Complete. Wednesday: Plate 1×10 6 cells ...cells at 3.8×10 6 cells per plate in DMEM Complete in 10 cm tissue culture plates. Incubate the cells at 37...previously seeded 10 cm plate. Slowly pipette the transfection mix onto the 10 cm plate, being careful not ...controller Hazardous waste container pH meter Stir plate Magnetic Stir Bar Reagents DMEM high glucose, Corning... in a T75 flask in 15 mL DMEM Complete. Friday: Plate 8×10 5 cells in a T75 flask in 15 mL DMEM Complete...

...week: Monday: Plate 1x10 6 cells in a 75 cm 2 flask in a volume of 15 mL. Wednesday: Plate 1x10 6 cells...cells at 3.8x10 6 cells per plate in DMEM complete in 10 cm tissue culture plates. Incubate the cells at 37... 10 cm plate). 56.7 µL of 1 mg/mL PEI, MW 25,000 Da in 443.3 µL of OptiPro SFM per 10 cm plate Pro-Tip...cells in a T75 flask in a volume of 15 mL. Friday: Plate 8x10 5 cells in a T75 flask in a volume of 15 mL...

...+ insert plate should have significantly more colonies than the recipient plasmid alone plate. The recipient...you can expect on your recipient plasmid + insert plate that are not correct. If you have a high number ...number of colonies on your recipient plasmid alone plate, you can try ligating the recipient plasmid alone ...transformation worked. You should also verify that you are plating on the appropriate antibiotic and try varying the...the number of background colonies on your control plate (the more background, the more colonies you will...

...+ insert plate should have significantly more colonies than the recipient plasmid alone plate. The recipient...If you are amplifying from a plasmid or simple template, there is very little chance for mis-priming, ...you can expect on your recipient plasmid + insert plate that are not correct. If you have a high number ...number of colonies on your recipient plasmid alone plate, you can try ligating the recipient plasmid alone ...the number of background colonies on your control plate (the more background, the more colonies you will...

...Microcentrifuge Electrophoresis chamber Power supply Rocking platform Fume hood Metal spatula Razor blade Plastic tray... in-house standard Before Starting Warm the hot plate to 100 °C. Thaw IgG standard and prestained protein...water for 5 min with gentle agitation on a rocking platform. Pour off the water in the sink. Add 20 mL of ...incubate for 1 h with gentle agitation on a rocking platform. Pour off the SimplyBlue SafeStain in the sink...incubate for 1 h with gentle agitation on a rocking platform. Pour off the water in the sink. Add 100 mL of...incubate for 1 h with gentle agitation on a rocking platform. Pour off the water in the sink. Take a brightfield...

...correspond to sequences on the template molecule (must be complementary to template strand). However, primers...to design primers that are complementary to the template region of DNA. They are synthesized chemically...primers do not need to correspond to the template strand completely; it is essential, however, that the 3’...end of the primer corresponds completely to the template DNA strand so elongation can proceed. Usually ...

...your LB agar plates are not fresh, you should streak your bacteria onto a new LB agar plate before growing... Streaking and Isolating Bacteria on an LB Agar Plate Creating Bacterial Glycerol Stocks for Long-term...commonly used to culture bacteria in the lab. LB agar plates are frequently used to isolate individual (clonal...toothpick, select a single colony from your LB agar plate . Drop the tip or toothpick into the liquid LB +...

...new window) Link to Video Making LB Agar Plates Create plates to culture bacteria in the lab Watch the...Video! Over-Agar Antibiotic Plating Quickly add antibiotic to a pre-poured plate Watch the Video! Streaking...Bacteria Isolate single bacterial colonies on an agar plate Watch the Video! Inoculating a Liquid Bacterial ...

...with shaking platform set to 120 rpm 37 °C bead bath Vortex Stir bar Magnetic stir plate pH meter Reagents...water in a 1 L bottle and stir on a magnetic stir plate. Stir until all particles have dissolved. Note: ...flask.Incubate in a 37 °C, 5% CO 2 incubator on a shaking platform set to 120 rpm. Pro-Tip Do not use cells that ...

...days. Single cells were then plated in individual wells of a 96-well plate and left undisturbed for 13 ... well (each well gets one dilution, so a 6-well plate will hold 5 dilutions plus one 'no virus' control...

...competent cells and retransform. Bacteria on an LB agar plate can be stored at 4°C for a few weeks. However, if...stock unthaw! Streak the bacteria onto an LB agar plate . Grow your bacteria overnight at the appropriate...

...(μL) Volume of Virus Added to Plate (μL) Volume of Cells Added to Plate (μL) Final Viral Dilution 1:10...

...controller Hazardous waste container pH meter Stir plate Magnetic stir bar Sonicator Ear protection Vortex...reach 100% confluence (80–90% is ideal). Pass and plate the cells the day before the transfection. Thaw ...

..., it is a good idea to place the gel on a glass plate if available. Unlike the plastic tray, this will...

...2-3μL into your favorite competent bacteria and plate. Be sure to pick multiple colonies for mini-prepping...

...sequence is used, plus a minimum of 18 bp of your template sequence. 5' and 3' primers will have different...

...liquid into different tubes or into wells of a microplate, you may consider using a multichannel pipette...

...bacterial culture. For details on how to streak a plate to get individual colonies and to generate liquid...