Molecular Biology: RNA, DNA, reverse transcription, exogenous spikes, & real-time PCR
Here is a product that does NOT work well with corals: Direct-zol (more to be added latter).
Here are protocols and reagents that DO work well:
1. RNA/DNA/protein extraction from endosymbiotic
anthozoans (corals and sea anemones) or see the book chapter
that outlines this method in more detail.
4 .TURBO DNA-Free (good when not using spin column kit)
5. High Capacity cDNA synthesis kit (a good, cheap, reverse
transcription kit from Life Technologies)
6. RNALater (works well for DNA & protein, but NOT RNA!)
7. Nucleic acids quantification spreadsheet (for "Nanodropping")
9. Budget and reagent checklist for real-time PCR experiments
10. EZ-TIME SYBR Green master mix brochure
Protein
19. 2D-Quant (for quantification of proteins in rehydration or SDS-PAGE buffer)
20. SYPRO Ruby (for staining SDS-PAGE and 2D gels)
21. Western blot of RBCL (can likely be used w/ other antibodies)
22. West Pico Chemiluminescent Substrate
23. Antibody for HSP70 (works for certain coral species)
24. HSP70 western blot with corals and Symbiodiniacieae
25. Ponceau staining (to check protein transfer to membrane)
26. Western blot trouble-shooting (from abcam)
27. Western blotting of cytoskeleton proteins (does not distinguish host from Symbiodiniaceae)
28. Antibodies that bind coral or Symbiodiniaceae antigens
29. Preparation of proteins for iTRAQ (label-based proteomics)
NMMBA's coral buoyant weighing station
Physiology
33. coral density calculations
35. Symbiodiniaceae density 36. surface area
I envision using 3D scanning tools for all future surface area measurements.
Blocks for coral surface area standard curve generation