MB Core Sample Requirements
For genomic DNA: Tail clips of .5 cm to 1 cm in length are preferred, placed one per well in the bottom of inserts in a 96 deep well culture plate. Alternatively, ear clips can be submitted, also in the bottom of inserts of a 96 deep well culture plate. It is most important that the tail/ear clips be of consistent size. There should be as little variation as possible across the samples submitted. If one clip is significantly
smaller than the rest, cut the rest to match it. We use a robotic procedure, and
consistent clip size will help ensure even DNA concentration of the final sample.
DNA should be in water. Moderately crude preparations are acceptable.
- Samples must be clean, submitted in water, and quantitated preferably using one of the Nanodrop instruments available on campus.
- Primers should be at a concentration of 3pmol/μl, and it is essential to provide a known annealing temperature or at least a Tm for the primer.
For automated sequencing on the ABI 3130 xL
- Plasmids – 400 ng of DNA in up to 12 μl water per reaction. A concentration of approximately 200ng/μl is ideal.
- PCR products –100 ng of DNA in up to 12 μl water per reaction for products up to 500 bp in size. Larger products will require more template (e.g., 200 ng for a 2000 bp product). Please include the size(s) of your products on your order.
Sample requirements for protein analyses will vary depending on the type of service requested. It is highly recommended that you consult with the MB Core Director to determine the best method for protein extraction and purification. In most cases, the MB Core will extract and purify proteins for analysis. The MB Core can also provide instruction and training on protein collection, as well as recipes for suitable protein extraction buffers.
Mammalian tissue: For protein analysis of mammalian tissue, it is recommended that the tissue be immediately flash frozen in liquid nitrogen after collection and transported to the lab on dry ice.
Cell cultures: For protein analysis of cell cultures, it is recommended that cell pellets be delivered on dry ice and and as much growth media is removed as possible.
Immunoprecipitation: For protein analysis following immunoprecipitation (IP), it is recommended that just the beads are submitted on wet ice at 4oC. Protein lysates submitted for analysis should
be free of contaminating charged molecules or ions like salts, ionic detergents, and DNA/RNA.
Protein storage: All samples should be kept @ -80oC to maintain high sample quality.