...FBS can be purchased already head inactivated or it can be inactivated in the lab by heating to 56 °C for...glutamine such as glutaGRO, Corning 25-015-CI) Heat-inactivated FBS Low serum medium such as Opti-MEM or Opti-Pro...bottle of DMEM high glucose, add 55 mL of heat-inactivated FBS and 5 mL of 100X glutaGRO. Store at 4 °C....

...recommended temperature should kill most potential contaminants, certain types of spores will stubbornly survive...For example: If you’ll be preparing plates with a final concentration of 100 ug/mL ampicillin, you should...growth of only the resistant strain. If you get this final result, your plates are ready for use or storage...any temperature and should always check for contamination prior to use. Negative Result 1: Both Strains...

...by restriction digest of existing sites in the original vector. Designing oligos To add NdeI, PacI, AscI... and CAGCT - 5' to the bottom oligo, making our final oligos 34 bp each: Top oligo: 5' - AATTC CATATG ...this would destroy the EcoRI and SalI sites in the final vector. Order the following oligos from your favorite...

... material you’re weighing and prevent cross-contamination with other substances. They also help you transfer...sterile (if necessary), to prevent any potential contamination. Scoop out the material you are weighing onto...much material, don’t put any extra back into the original container. Dispose of the excess material into...

...Background Information The final step in the construction of a recombinant plasmid is connecting the insert...vector Insert or water + Any colonies indicate contamination of intact plasmid in ligation or transformation...

...DNA segments you will need to join to create your final plasmid. Adjacent segments should have identical...digest . Sequence the important regions of your final plasmid, particularly the seams between the assembled...especially useful for introducing promoters, terminators, and other short sequences into the assembly ...

...where the image is further magnified before it finally reaches your eyes. These lenses determine the magnification...lens provides 10x magnification. To determine the final magnification of your image, multiply the magnification...sample. To ensure that your sample is adequately illuminated, look through the eyepiece. Adjust the power ...

...Repeat steps 2-4 25-30 times. Final Extension for 5 minutes at 72°C: A final extension to fill-in any protruding...minutes at 72°C. Repeat steps 8-10 for 25-30 cycles. Final Extension for 5 minutes at 72°C. Run 2 μL on a gel...

...band to cut without having DNA in other lanes contaminating your sample. To accomplish this, it is best ...each buffer to add during the DNA isolation step. Finally, you will want to isolate the DNA from the gel....

...produce antibodies Antibody Purification Purify recombinant antibodies using Protein A or Protein G columns... Stain Determine purity and concentration of recombinant antibodies Watch the Video! Western Blot Separate...

...of the buffer will evaporate and thus alter the final percentage of agarose in the gel. Many people prefer...mins. (Optional) Add ethidium bromide (EtBr) to a final concentration of approximately 0.2-0.5 μg/mL (usually...

...higher competency cells. Additionally, if your final product is going to be very large (>10kb) you might...ligation reaction. Isolate the Finished Plasmid Finally, you will need to pick individual bacterial colonies...

...Antibody Blog Posts Immunocytochemistry Protocol Recombinant Antibody Purification Protocol Molecular Biology...Assay Kit or other preferred method for protein determination. Prepare a 50:1 Reagent A to Reagent B dilution...

...equipment, and reagents RNase-free (Use an RNase decontamination solution, such as RNaseZap® or RNase AWAY®,...microcentrifuge tubes, 4 mL polypropylene tubes RNase decontamination solution like RNase AWAY® or RNaseZAP® Isopropanol...

... primers) or too short. Short primers produce inaccurate, nonspecific DNA amplification product, and long...

...addgene.org and include: The sequencing result The original trace file (.ab1 file) provided by your sequencing...

...will not be gel purifying it, you may need to inactivate the enzyme(s) following the digestion reaction...

...Collection Antibody Blog Posts Western Blot Protocol Recombinant Antibody Purification Protocol Molecular Biology...