...Bacterial Glycerol Stocks for Long-term Storage of Plasmids Creating Bacterial Glycerol Stocks for Long-term...Bacterial glycerol stocks are important for long-term storage of plasmids. Although you can store your plasmid...for creating a glycerol stock, including additional tips for long-term storage. Protocols...LB agar plate can be stored at 4°C for a few weeks. However, if you want to store bacteria for a longer...keeping the cells alive. A glycerol stock of bacteria can be stored stably at -80°C for many years. Protocol...unexpectedly at -80°C. Freeze the glycerol stock tube at -80°C. The stock is now stable for years, as long as...concentration of long-term glycerol storage is unknown. Most labs store bacteria in 15-25% glycerol. You ...

...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...

...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...

...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...

... 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...

... 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....

... 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...

...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...

...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...

...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...

... 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....

...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...

...into sterile tubes. Store the tubes at -80 °C. After thawing, the solution can be stored at 4 °C for up to... glutaGRO 11 mL of 200 mM L-alanyl-L-glutamine. Store at 4 °C. D1 + 0.1 M sorbitol (optional) : DMEM ...months, discard the tube and thaw a new working stock. 40% POLYETHYLENE GLYCOL (PEG) 8000 solution: Dissolve... solution may separate into phases. Aliquot and store at 4 °C. Cell Lysis Buffer : 50 mM Tris HCl, 150....5 Filter sterilize through a 0.22 μm membrane. Store at 4 °C. Considerations Before You Start The health...

...Agar Plate Creating Bacterial Glycerol Stocks for Long-term Storage of Plasmids Recovering Plasmid DNA from... For long term storage of the bacteria, you can proceed with Creating a Glycerol Stock . You can now isolate...temperature. Now screw on the top of the bottle and store the LB at room temperature. When ready to grow your...

...Prepare 25 ng/µL stocks of each sample. Samples can be used for ddPCR immediately or stored at -20 °C until...For the first virus dilution, add 160 µL of viral stock to 240 µL of media. When adding the virus to the...Dilute Media Volume Total Volume 2.5 160 µL viral stock 240 µL 400 µL 5 200 µL of 2.5-fold dilution 200 ... is an additional 10-fold dilution of the virus stock! Don’t forget to factor this into your calculations...transduce cells: Initial dilution factor of virus stock Dilution factor after additional 10-fold dilution...

...much virus as possible. Store at 4 °C for short term (2 weeks), or aliquot and store at -80 °C for long term...Sterilize by passing through a 0.22 μm filter and store at 4 °C. 1X PBS-MK buffer Dissolve 26.3 mg of MgCl...Sterilize by passing through a 0.22 μm filter and store at 4 °C. 0.1% Pluronic-F68 in PBS (A) Add 500 µL...clean fractions. Concentration and buffer exchange: Stocks of Pluronic F68 Pluronic F68, 10% solution A: 0.1%...

... mM EDTA Store Solution I at 4°C Solution II - Denaturing Solution 0.2 N NaOH 1.0% SDS Store Solution ...acetate 23 mL glacial acetic acid 57 mL of dH 2 O Store Solution III at 4°C Grow 2 mL overnight cultures...containing your plasmid of interest. Add 1.5 mL of the stock culture to a 1.75 mL microfuge tube. Centrifuge ... room temperature before quantifying and using. Store DNA at 4°C. Tips and FAQ Plasmid purification kits...

...Bacterial Culture Introduction If you have a glycerol stock or stab culture of bacteria and need to purify plasmid...population (single colony) of bacteria from this stock. Using a single colony from a freshly streaked agar...Obtain the approrpriate bacterial stab or glycerol stock . Using a sterile loop, pipette tip or toothpick... of the stab culture or the top of the glycerol stock. Pro-Tip If you use a wire loop you can sterilize...

...viral stock will be higher titer, consider additional dilutions. Dilution Volume of Lentivirus Stock (μL...heat-inactivated FBS and 5 mL of 100X glutaGRO. Store at 4 °C. Pro-Tips Different brands and lots of FBS...it is recommended that you titer from the frozen stock to account for any loss in titer associated with...

...Reagents TAE ( recipe here ) Agarose Ethidum bromide (stock concentration of 10 mg/mL) Procedure Pouring a Standard...Pro-Tip It is a good idea to microwave for 30-45 sec, stop and swirl, and then continue towards a boil. Keep...approximately 0.2-0.5 μg/mL (usually about 2-3 μl of lab stock solution per 100 mL gel). EtBr binds to the DNA ...sample is unloaded, push the pipettor to the second stop and carefully raise the pipette straight out of ...