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TypeProtocol...sample (do not vortex) Incubate 30 min at 37 °C Transfer to ice ** Critical: do NOT treat your plasmid ...dilutions 5–8 Note: at Addgene, samples typically range from 1 x 10 12 GC/mL to >2 x 10 13 GC/mL and we ... and forth at least 5 times. Seal plate with transparent film. Centrifuge at 3,000 rpm for 2 min to bring...) ~ 1.0, E (efficiency of PCR) ~100% (90%–110% range is acceptable) Baseline removal: all samples will...
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Affinity Purification of Recombinant Antibodies with Protein A or Protein G
TypeProtocol...Collection Antibody Blog Posts Antibody Guide Transfection for Recombinant Antibodies Immunocytochemistry... Inclusion of this information is solely for transparency intended to support reproducibility in science...recombinant antibody to room temperature (see Transfection for Recombinant Antibody protocol ). Pro-Tip... has reached the desired concentration. Gently transfer the sample from the reservoir to a LoBind tube... -
AAV ddPCR Titration
TypeProtocol...outlined in this protocol are based on an AAV titer range of 5E+12–5E+13 GC/mL, where GC is genome copies ...the DG8 gasket, making sure that it is secure. Transfer the cartridge holder to the droplet generator....while slowly and gently aspirating the droplets. Transfer the droplets to a prechilled PCR plate. Pro-Tip... 98 10 2 1 Hold 4 ∞ 2 1 After PCR is complete, transfer the plate to the Droplet Reader. Open the QuantaSoft... -
Protocol - How to Design Primers
TypeProtocol...complementary to template strand). However, primers do not need to correspond to the template strand completely; it...primer corresponds completely to the template DNA strand so elongation can proceed. Usually a guanine or... -
Protocol - How to Create a Bacterial Glycerol Stock
TypeProtocol... of Plasmids You may also like... Bacterial Transformation Recovering Plasmid DNA from Bacterial Culture... not need to obtain more competent cells and retransform. Bacteria on an LB agar plate can be stored at...frozen bacteria, preventing damage to the cell membranes and keeping the cells alive. A glycerol stock ... -
Antibody Validation Using the Indirect ELISA Method
TypeProtocol... Inclusion of this information is solely for transparency intended to support reproducibility in science...case, use lower antibody concentrations. The ideal range of the standard curve will vary between targets ...your unknown sample’s absorbance falls above the range of the standard curve you will need to either include... -
Kit Free RNA Extraction
TypeProtocol...cells. Extract RNA from the homogenized sample(s).Transfer tissue/cell lysate to a 4 mL tube. Add the following...the cold room may damage it. Using a pipettor, transfer the top, aqueous phase to a new RNAse-free tube... -
DNA Quantification
TypeProtocol...spectrophotometer. A spectrophotometer uses the absorbance/transmission of light through a liquid to determine the concentration...measured under the 260/280 column (A good purity ranges from 1.80-2.00). Repeat for each sample. Notes:... -
Plasmid Modification by Annealed Oligo Cloning (with Protocols)
TypeProtocol...Digest of Plasmid DNA DNA Ligation Bacterial Transformation Summary Oligo overlap cloning can be used anytime...the vector with 0.75-6 ng of annealed oligos). Transform 2-3μL into your favorite competent bacteria and... -
Using a Light Microscope Protocol
TypeProtocol...your cell culture dish. Microscopes come in a huge range of shapes and sizes - from phone-sized, foldable...powerful (but are cheap and accessible) to massive transmission electron microscopes that allow us to see cellular... -
Protocol - How to Run an Agarose Gel
TypeProtocol...the solution has a tendancy to boil over. Placing saran wrap over the top of the flask can help with this...are usually referred to as ‘bands’ due to their appearance on the gel. Pro-Tip If you will be purifying ... -
Pouring LB Agar Plates
TypeProtocol...x (0.220 L) = 8.14 g pre-mixed LB-agar powder. Transfer the LB-agar powder you’ve measured out into an...molten agar from boiling over in the autoclave. Transfer the sterile water (in our case 220 mL) to the ... -
CRISPR Guide
TypeGuide..., 1163–1171. PMID: 23979020 Cong, L., Ran, F. A., Cox, D., Lin, S., Barretto, R., Habib, N., Hsu, P. D...38688943 Hsu, P. D., Scott, D. A., Weinstein, J. A., Ran, F. A., Konermann, S., Agarwala, V., Li, Y., Fine..., 17 (5), 471–479. PMID: 32203383 Nishimasu, H., Ran, F. A., Hsu, P. D., Konermann, S., Shehata, S. I....CRISPR-Cas9. Cell , 167 (7), 1829-1838.e9. PMID: 27984730 Ran, F. A., Hsu, P. D., Wright, J., Agarwala, V., Scott...Nature Protocols , 8 (11), 2281–2308. PMID: 24157548 Ran, F. A., Cong, L., Yan, W. X., Scott, D. A., Gootenberg...may be a single-stranded oligonucleotide, double-stranded linear DNA, or a double-stranded DNA plasmid. ...including transposons, integrases, and recombinases, with Cas enzymes. CRISPR Transposases Transposon systems... -
Protocol - How to Streak a Plate
TypeProtocol...also like... Making LB Agar Plates Bacterial Transformation Recovering Plasmid DNA from Bacterial Culture... -
Protocol - How to Purify DNA from an Agarose Gel
TypeProtocol... gels, so you will want to stay in the 0.7-0.8% range if possible. You will want nice crisp bands. This... -
Protocol - How to Perform a Diagnostic Digest
TypeProtocol... bacterial endonucleases that recognize a large range of DNA sequences. Given the variety of these enzymes... -
Centrifugation
TypeProtocol... tubes. Depending on the centrifuge their speed ranges can vary, but are typically lower than some other... -
Molecular Biology Protocol - Restriction Digest of Plasmid DNA
TypeProtocol...amount of DNA in the reaction, incubation time can range from 45 mins to overnight. For diagnostic digests... -
Pipetting Protocol
TypeProtocol...boxes that the tips come in often indicate a volume range that the tip can hold. This should give you an idea... -
AAV Purification by Iodixanol Gradient Ultracentrifugation
TypeProtocol... ml B + 200 mM NaCl Procedure Cover the filter membrane with 15 mL of 0.1% Poloxamer 188 PBS and incubate...