We narrowed to 728 results for: STO;
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TypeGuide...VSV-G due to wide infectivity VSV-G (vesicular stomatitis virus G protein) is the most common envelope ...supernatant containing the viral particles is removed and stored or centrifuged to concentrate. Crude or concentrated... The viral envelope gene; typically vesicular stomatitis virus G glycoprotein (VSV-G), an envelope protein...
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Plan Your Experiment
TypeGuide...using CRISPR . Browse Addgene's collection of in-stock lentiviral preps . AAV vectors AAV vectors offer...overcoming the AAV size limitation . Browse Addgene's in-stock AAV preps or try Addgene's Packaged on Request service...paired with restriction enzymes for further customization Usually paired with a sequencing method Can’... -
CRISPR Guide
TypeGuide...insertions, or frameshift mutations leading to premature stop codons within the open reading frame (ORF) of the... cytosines in a gene’s promoter or by inducing histone acetylation or demethylation. The most common modifiers... green fluorescent protein (GFP), creating a customizable DNA or RNA label for fluorescence microscopy...gene by shifting the ORF and/or creating premature stop codons NHEJ N on- H omologous E nd J oining; A DNA... -
CRISPR/Cas9 FAQs Answered!
TypeBlog Post...transfection, you could use 1 µl of 10 µM oligo stock for each well on 24-well plate, co-deliver into ... -
CRISPR 101: Cytosine and Adenine Base Editors
TypeBlog Post...from Nicole Gaudelli in David Liu’s lab in 2016. History of base transition editing Cytosine base editing... -
CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More
TypeBlog Post... due to its condensed design. For both systems, custom spacers/gRNAs are easily inserted using BbsI and... -
Troubleshooting and Optimizing a Western Blot
TypeBlog Post...accurate Check plasmid sequence for frameshifts and stop codons Weird background patterns (dots, lines... -
Adeno-associated virus (AAV) Guide
TypeGuide...capsids for more information. Browse Addgene’s in-stock viral preps . AAV retrograde (AAVrg) Directed evolution...Packaging Services Packaged-on-Request AAV Preps In-Stock AAV Preps Resources on addgene.org AAV Plasmids ... -
22 Hot Plasmid Technologies from 2014
TypeBlog Post...family of GFP proteins in the cephalochordate Branchiostoma floridae, named bfloGFPs. They went on to characterize... -
Antibody Validation Using the Indirect ELISA Method
TypeProtocol...ng/µL stock into 900 µL PBS in a microfuge tube and vortex. 1 ng/µL : Add 450 µL of 2 ng/µL stock into ...multichannel pipette, add 100 µL of TMB Stop Solution directly to the wells to stop the reaction. Measure the absorbance...different incubation options available. Caution Use the Stop Solution in a well ventilated area and avoid breathing...tube and vortex. 0.5 ng/µL : Add 450 µL of 1 ng/µL stock into 450 µL PBS in a microfuge tube and vortex. .... 0.25 ng/µL : Add 450 µL of 0.5 ng/µL stock into 450 µL PBS in a microfuge tube and vortex. 0.125 ng/...ng/µL : Add 450 µL of 0.25 ng/µL stock into 450 µL PBS in a microfuge tube and vortex. 0 ng/µL : 450 µL...concentrations of HRP yield a greenish solution. Stop the reaction before any wells display a green product... -
Lentivirus Production
TypeProtocol...sterile tubes. Store the tubes at -20 °C or -80 °C. After thawing, the solution can be stored at 4 °C for... the harvested media to a polypropylene storage tube and store at 4 °C between harvest. Centrifuge the... of heat-inactivated FBS and 5 mL of glutaGRO. Store at 4 °C. Pro-Tip Different brands and lots of FBS...months, discard the tube and thaw a new working stock. The optimal mass DNA:mass PEI ratio will need to... to batch variation when making the PEI working stock, therefore the ratio of μg DNA:μg PEI needs to be....45 μm PES filter. The viral supernatant can be stored at 4 °C for several hours but should be aliquoted...aliquoted and snap frozen in liquid nitrogen and stored at -80 °C as soon as possible to avoid loss of titer... -
Pipetting Protocol
TypeProtocol...multiple “stops” on the plunger, where you feel resistance while pressing down on the pipette. Stop at the...the first “stop.” The first stop expels enough air from the pipette for an accurate measurement in the ...way to the second stop to release the liquid. Pro-Tip Dispense until the first stop, wait, and then dispense...the next step. The second stop is meant to expel excess liquid from the tip once you’ve reached step 8. ... to its resting position. Wait for the liquid to stop flowing into the pipette tip. Liquids of various...dispense to the second stop to expel any residual liquid in the tip. Before releasing the plunger, lift... -
What is Polymerase Chain Reaction (PCR)
TypeProtocol... 2.5 μL Forward Primer (10 μM stock) 2.5 μL Reverse Primer (10 μM stock) 36.8 μL Sterile dH 2 O 0.2 μL... 2.5 μL Forward Primer (10 μM stock) 2.5 μL Reverse Primer (10 μM stock) 0.2 μL Taq DNA Polymerase (5 ...making a 1:10 dilution of the stock. For example, add 100µl of primer stock to 900µl of dH 2 O. Tips and...liquid, normally sterile dH 2 O. To make a 100uM stock of any primer, add a number of µl of dH 2 O equal...38.5nm, add 385µl of water. After making your 100uM stock, immediately make a working concentration of each...to supplement the amount supplied by the buffer stock, where it is known to form a gradient when frozen... -
Transfection for Recombinant Antibodies
TypeProtocol... A6279 Before Starting Warm the DNA and working stock of PEI-MAX to room temperature before use, and warm...sterile 0.22 µm filter. Prepare 10 mL aliquots and store the solution at -20 °C. 1 mg/mL PEI-MAX Add 1 g ... through a 0.22 µm filter. Prepare aliquots and store at -20 °C until use. BCD TFX 1000 mL BalanCD HEK293...mL 200 mM Glutagro Do not add selective reagents Store at 4 °C until use. We suggest preparing fresh solutions...500 mL BalanCD HEK293 Feed 20 mL 200 mM Glutagro Store at 4 °C until use. We suggest preparing fresh solutions... the second tube of 6 mL BCD TFX (for a 1 mg/mL stock solution of PEI-MAX, 450 µg = 450 µL). Cap the tube...our Recombinant Antibody Purification Protocol ). Store the supernatant at 4 °C until ready to use.... -
AAV Titration by qPCR Using SYBR Green Technology
TypeProtocol... plates) and store at -20 °C. Once a standard is thawed do not freeze it again but store at 4 °C and use...standard curve plasmid (2 x 10 9 stock made in step #1): Volume of 2 x 10 9 stock or previous dilution (μL) ...Water 4.7 μL 470 μL Procedure Prepare a plasmid stock of 2 x 10 9 molecules/μL to generate a standard ...79.8 = 1.25 μL Therefore we need to dilute 1.25 μL stock into 98.74 μL H 2 0 Pro-Tip Once a validated standard...Total dilution Dilution 1 (DNase step) 5 uL AAV stock 45 uL 10X 10X Dilution 2 5 uL Dil. 1 95 uL 20X 200X... -
Pouring LB Agar Plates
TypeProtocol...your plates are ready for use or storage at 4 ℃. You should not store your plates for longer than 1 month...Antibiotic Concentrations Antibiotic Recommended Stock Concentration Recommended Working Concentration ...to the molten gel mix, you should create a 1000x stock solution. For example: If you’ll be preparing plates...concentration of 100 ug/mL ampicillin, you should make a stock solution of 100,000 ug/mL (100 mg/mL). Simply measure...solidifies in the bottle while you’re pouring, you should stop pouring and re-make the gel-mix. If you’re making...material to reduce condensation. The plates are then stored at 4 ℃ until use. Once your plates have solidified... -
Protocol - pLKO.1 – TRC Cloning Vector
TypeProtocol...to a polypropylene storage tube. The media contains your lentiviral particles. Store at 4°C. m. Add 5 mL...may be stored at 4°C for a few days, but should be frozen at -20°C or -80°C for long-term storage. TIP:...concentration of 100 μg/mL antibiotic. Pour plates and store at 4°C. Hexadimethrine Bromide (Polybrene) Prepare...catalog #H9268) in 0.9% NaCl. Autoclave to sterilize. Stock solution is stable at 4°C for up to one year. The...stable at 4°C for several years. Protamine Sulfate Store protamine sulfate (MP Biomedicals catalog #194729...soluble in cold water. Puromycin Prepare a 50mg/mL stock solution of puromycin (Sigma-Aldrich catalog #P8833... Sterilize by passing through a 0.22 μm filter. Store aliquots at -20°C. I.3. Warranty Information Addgene... -
General Transfection
TypeProtocol...filter. Aliquot 50–100 µL and store at -20 °C. Aliquots can be thawed and stored at 4 °C prior to use. Thawed...into sterile tubes. Store the tubes at -80 °C. After thawing the solution can be stored at 4 °C for up to...heat-inactivated FBS and 5 mL of 100X glutaGRO. Store at 4 °C. Pro-Tip Different brands and lots of FBS...months, discard the tube and thaw a new working stock. Considerations Before You Start The health of the... -
Affinity Purification of Recombinant Antibodies with Protein A or Protein G
TypeProtocol... sterile sodium phosphate dibasic (NaH 2 PO 4 ) Store up to 1 month at 4 °C Adjust pH to 7.0 Autoclave...Gravitrap columns and gently pour off the ethanol storage buffer. Tightly attach LabMate PD10 Buffer Reservoirs...washing with 25 mL of Protein A/G binding buffer and store in 20% ethanol at 4 °C . Pro-Tip Columns may be ...Centrifuge column at 1000 x g for 2 min to remove storage solution. Place a mark on the side of the column...antibody to 1 mg/mL with PBS if needed. For long term storage, add sterile sodium azide to 1 mM. Option 2: Buffer... the NanoDrop Spectrophotometer. For long term storage, add sterile sodium azide to 1 mM.... -
Kit Free RNA Extraction
TypeProtocol...equipment back to room temperature, as prolonged storage in the cold room may damage it. Using a pipettor...quantification , which can be modified for RNA. Store your RNA sample(s) at -80 °C to prevent RNA degradation...sample, consider making smaller aliquots of it and storing those in -80°C. Option #2 - TRIzol® Protocol Homogenize...DNA quantification , which can be modified for RNA.Store your RNA sample(s) at -80 °C to prevent RNA degradation...sample, consider making smaller aliquots of it and storing those in -80°C. Solution D Protocol TRIzol® Protocol...