The Problem with Molecular Biology Techniques
I taught myself how to program a computer. It took a long time to become a fluent programmer, but there are a fair number of good
books on most programming languages. Learning is a matter of patience, perserverance, and time spent in front of the computer.
When I began learning molecular biology techniques (e.g. cloning, sequencing, PCR, etc...), I disliked how much you have to
rely on other people to show you things and explain things to figure out the different techniques - there isn't a molecular biology
compiler that will print an error message saying, "warning your dNTPs have gone bad, please retry your PCR with freshly prepared
dNTPs". It feels like you're constantly bothering other people. And if you ask two different people how to make a particular buffer
or run a certain reaction, you'll get two slightly different methods - making it hard to know exactly what you should do.
There have been a few books more useful than others and a few mistakes that I now know how to avoid,
so I put this kinda thing here when I think of it.
|something won't go into solution|
| ||Is the pH right? many things (e.g. EDTA!!!) won't go into solution until they are at a certain pH|
| ||try heating it up a bit|
| ||did you add too much? (you can check percent solubility on some MSDS sheets)|
|a protocol for TRIS says to add HCl until you reach a certain pH, but after adding TRIS you find the pH already too low.|
| ||there are two kinds of TRIS 1=Tris Hydroxymethyl Aminomethane Hydrocloride and 2=Tris Hyroxymethyl Aminomethane.
You used 1, but your protocol used 2.|
Tricks to make life easier
||glycerol is commonly used in freezer stocks (300uL in 2mL total volume freezer stock),
but its high viscosity makes it difficult to pipette. Typically glycerol is autoclaved to sterilize it. After the glycerol has cooled
enough that you can touch the bottle without burning yourself, it is a good time to aliquot it to sterile freezer stock tubes, because hot
glycerol pipettes like water.