Post date: Oct 15, 2017 6:12:47 PM
Filling Jars
1) There shalt not be any open containers of beans in lab.
2) Each 1/2 gallon ball jar should contain 2000 beans: fill to the 1000-mark/2-mark on the jar
3) Cap jar with lid or mesh
Isolating Beans
1) Wait to isolate until the first few beetles start to emerge from the standard/stock colony.
-This maximizes synchronization of hatch, and limits the amount of time spent in pill capsule.
2) Sieve beans from stock colony and use adult-free beans to fill pill capules
3) Place a single bean in each pill cap to reach 2000/population (6000 beans total; 1000 from each of the six stock jars)
4) Store pill caps in incubator; remove emergers daily
Making New Colonies
1) When enough adults are present, sieve emergers (use beans from BOTH stock colony jars)
2) Add ~2-3000 beetles to new jar (determine by volume: see line marked on bottom of sieve)
3) After 8-10 days, sieve off all adults (to avoid excess larvae without decreasing Ne (as much), sieve adults early??)
-Too many beetles/bean = excessive metabolic heat + humidity = mold + death + bitter recrimination
Making Controls
1) Chill jars for 24 hours to standardize with the chilled experimental beetles
2) Sieve emergers (use beans from BOTH stock colony jars)
3) Weigh 3.3 g of beetles (approximately 1000 beetles) to found the control colony
a) pour beans into a large petri dish
b) freeze the petri dish for a few minutes/until beetles stop crawling
c) shake petri dish to allow beetles to fall to the bottom and skim off beans by slowly pouring them into a new dish
d) pour beetles into the weigh boat (with lid!)
e) repeat until 3.3 g of beetles are obtained (fills a 10-mm petri dish to a depth of approximately 2-3 beetles)
4) Add to a jar of 2000 beans
mass.50.beetles.in.g <- c(0.15219, 0.14915, 0.15464, 0.17766, 0.17109, 0.19123, 0.15229, 0.15337, 0.17369, 0.18664, 0.16755, 0.18127, 0.15034, 0.17948, 0.13848)
ave. mass 1000 beetles = 3.305427 g +/- ~0.3 g
Sexing Virgin Beetles
1) First person sorts pill caps into containers of emerged beetles vs. no emerged beetles
2) Second person sorts virgin beetles to reach 250 of each sex for each population (isolate 1500 total per day)
a) Virgin beetles are defined as 1) the single emerger in a pill capsule or 2) emergers of a single sex in a pill capsule
b) Track number of virgin beetles added to a dish using tally marks
c) Non-virgin beetles should be disposed of
d) Chill petri dishes with 250 virgin beetles in fridge at 10 C overnight
e) Repeat the following day to reach 3000 total virgin beetles (in petri dishes of 250 each)