...deposit, associate the plasmids with a Principal Investigator (PI), provide us with your shipping address ...

...bacterial colonies. Equipment Shaking incubator at 37 °C Stationary incubator at 37 °C Water bath at 42 °C Ice...temperature and then (optional) incubate in 37°C incubator. Mix 1 - 5 μl of DNA (usually 10 pg - 100 ng) ...antibiotic) to the bacteria and grow in 37°C shaking incubator for 45 min. Pro-Tip This outgrowth step allows... do this you will need to have access to an electroporator and the appropriate cuvettes. Follow the manufacturer's...

...19246619.Downey N. Interpreting melt curves: An indicator, not a diagnosis. IDT, 2014. Dwight Z; Palais ...

...personal protective equipment including laboratory coat, laboratory goggles, and gloves. Sodium azide containing...Class II, Type A2 Biological Safety Cabinet 4 °C Refrigerator Pipette controller Benchtop centrifuge compatible...tubes NanoDrop spectrophotometer 37 °C, 5% CO 2 incubator with shaking platform set to 120 rpm 37 °C bead...upright at -20 °C Procedure Section 1: Affinity chromatography Equilibrate filtered tissue culture supernatant...

...addressed by the design of the vectors and by safe laboratory practice. In terms of vector design, 2nd and ...Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element Enhances Expression of Transgenes Delivered... JM, Hughes SH, Varmus HE. Cold Spring Harbor Laboratory Press. Cold Spring Harbor, 1997. Available online...export. Tat in trans Second generation only; Trans-activator; binds TAR to activate transcription from the ...Woodchuck hepatitis virus post‐transcriptional regulatory element; sequence that stimulates the expression...

...Trypanosoma cruzi yes, cut S. pyogenes Neo Docampo tdTomato/pTREX-b 68709 Other/Trypanosoma cruzi none S. ...

..., Zalatan JG (2018) Synthetic CRISPR-Cas gene activators for transcriptional reprogramming in bacteria...

...can be replicated easily and efficiently in a laboratory setting. Stable - Plasmids are stable long-term... contain a promoter sequence, a transcription terminator sequence, and the inserted gene. The promoter...RNA from the insert DNA via transcription. The terminator sequence on the newly synthesized RNA signals...design a effective primer. Sanger sequencing chromatogram Sanger sequencing is modeled after in vitro ...sequencing results are presented as a sequencing chromatogram which provides the color and intensity of each...

...Harvest antibody Equipment Biosafety Cabinet 4 °C Refrigerator Micropipettes and tips Pipettor and pipettes...conical tubes Automated cell counter 37 °C, 5% CO 2 incubator with shaking platform set to 120 rpm 37 °C bead... mL vented flask.Incubate in a 37 °C, 5% CO 2 incubator on a shaking platform set to 120 rpm. Pro-Tip ...swirl 5–10 times to mix. Return the flask to the incubator. Section 3: BCD Feed and valproic acid supplementation...

... the two, use a conversion calculator or chart like the G Force calculator (Link opens in a new window...Introduction A centrifuge is a common piece of laboratory equipment used to separate different components...

...water Microscope slide Anti-fade mounting medium Laboratory wipes 15 mL conical tubes 50 mL conical tubes...paraformaldehyde and follow your institution's laboratory safety guidelines for disposing of waste in the...remove the coverslip. Blot the coverslip with a laboratory wipe to remove excess liquid. Add 1 drop of anti-fade...

...without permanently modifying the genome dCas9-activator (such as dCas9-VP64) gRNA(s) targeting promoter...promoter elements of target gene Many different activators exist, including the multi-plasmid SAM system. Select...activate or repress a target gene using dCas9-activators or dCas9-repressors, gRNAs should be targeted...

...pipette 200–1000 µL single channel pipette CO 2 incubator Pipet controller Hazardous waste container Microscope...orange/yellow if using a standard phenol red pH indicator) because the buildup of waste products could be...difficult to find. Leave the cells undisturbed in an incubator for 7–14 days. After ~7 days, scan the plate for...

...require modifications dictated by the originating laboratory for optimal results. If you obtained the pooled...limited. Equipment Table top centrifuge BioRad Electroporator (MicroPulser TM , Bio-Rad 1652100) Reagents...time for a total of eight electroporations): Electroporator Conditions: Bio-Rad Micropulser Ec1 0.1 cm ...

...and bathroom. These items should remain in the laboratory. When traveling between lab spaces, take your...adhered to. None of the authors, contributors, administrators, or anyone else associated with Addgene, can...

...loop Bunsen burner (or other small flame source) Incubator Marker Reagents LB agar plate (with appropriate...resistance and your initials. Labeling within a laboratory setting is important for organization, and it...

...spreading of antibiotic makes it easy for an investigator to conveniently plate and select transformed...spreading Bunsen burner (or other small flame source) Incubator Reagents 6 cm diameter LB/agar plates without ...

...Addgene’s Mammalian RNAi Tools page. Several other laboratories have deposited pLKO derived vectors that may... temperature. g. Retrieve HEK-293T cells from incubator. The cells should be 50-80% confluent and in DMEM...should be decontaminated with at least 10% bleach. Laboratory materials that come in contact with viral particles...

... lab! Introduction A water bath is a piece of laboratory equipment that helps bring your materials to .... Water baths can also be placed in walk-in refrigerators to achieve a temperature between 4°C and room...

... Protocols Video protocol guides for standard laboratory procedures. How-To Videos Screencapture walk-...Electrophoresis Gel electrophoresis is the standard laboratory procedure for separating DNA by size for visualization...