Written high-level notes that might help you decide on directions. Basically most bio experiments require the following five steps. Have spelled out the first stage. If you are interested, I can expand on the methods and materials required for all five stages.
- Grow - microbial cells, plant cells, animal cells.
- Sources of materials:
- Microbial cells: Brewers yeast, Curd yogurt for lactobacilli, feces for E.coli, Probiotic capsules (various)
- Plant cells: Any small weeds (oxalis/wood sorrel is a great examples), algae from ponds
- Animal cells: Protozoa from stagnant water (amoeba, paramecium, tetrahymena). Worms, insects. Fresh muscle cells from food meats (poultry, beef) for fibroblasts.
- Whole organisms
- Equipment:
- Incubators for growing cells (DIY options)
- Blenders/homogenizers to break tissues into cells (Kitchen supplies)
- Colorimeters to measure cell densities (DIY options)
- Microscope (DIY or used options)
- Consumables:
- Growth medium to keep cells/tissues happy (bacterial cells in protein and yeast extract powders from health stores, same for yeast and most protozoa; hydroponics materials for plants)
- Containers for medium and other materials
- Measuring stuff. Measuring spoons, cups, pipettes. Weighing scale
- Purify
- DNA
- Method 1: Grind, digest with proteases (contact lens cleaners, meat tenderizer tablets) and dissolve with detergents. Add some kind of salt, add alcohol. DNA precipitates out. Spool or centrifuge the DNA. Dissolve in buffer containing EDTA (Versene). EDTA blocks enzymes that digest DNA.
- Method 2: Grind, digest and dissolve as in method 1. Add Silica or Alumina powders (check sand blasting suppliers). These powders bind to DNA preferentially at high salt. Remove powders with bound DNA by centrifugation. Wash once with high salt buffer. Release DNA with low salt buffer. Under some conditions DNA binds more tightly to alumina.
- Method 3: Grind, digest and dissolve as in method 1. Add beads that have pores in them to hold back small molecules (Sephadex G50, etc.). The junk gets held back (amino acids, peptides, salts, small metabolites) and the large molecules like DNA and RNA are excluded. Remove the beads by centrifugation or filtration and the liquid (supernantant) will contain the crude nucleic acids.
- RNA
- Similar methods to purify as above but RNA is very labile, breaking down easily because of contaminating enzymes that digest RNA that are pretty much everywhere. Equipment used in RNA isolation needs to be pre-processed to be RNAse free. There are tricks to convert RNA directly into DNA and then doing some of the analysis on the cDNA made from the RNA. The steps are not difficult.
- Proteins
- Macromolecular complexes
- Metabolites
- Manipulate
- Analyze
On Sat, Sep 17, 2016 at 12:26 PM, 'Tobias' via DIYbio <diybio@googlegroups.com> wrote:
I am not sure about that yet. What is less expensvie?--
I have another question. Is it allowed in Germany to do Biohacking?I readed that in the EU only Labours with permission are allowed to perform such actions.
Am Freitag, 16. September 2016 20:18:26 UTC+2 schrieb Tobias:HelloI recently found the topic Biohacking and think its really amazing.Now i want to start on my one. The Question is what kind of Equipment I need.I already startet reading and looing videos about gentik.Can someone suggest some general instruments and machines i need?Mfg Tobias
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