Team:Purdue/Seed plating

(Difference between revisions)

Revision as of 16:23, 8 July 2010

sterile seed in deluted agar
sterile agar media
sterile hood environment
P1000 pipeter
blue pipeter tips
plates
parafilm tape
70% ethanol solution in spray bottle
grid template for desired seed alignment (to be placed under transparent media and referenced during seed placement)

Pouring Agar

sterilize items one by one with 70% ethanol spray, placing each item into sterile hood after being sprayed.
once all are in hood, remove aluminum foil wrap from agar flask.
remove lid off first plate and keeping the lid as a shield between you and the open plate, pour agar into plate. Fill to halfway point, carefully avoiding air bubbles in media. Place lid right-side-up on hood surface, then set poured plate on top. Allow to cool before replacing lid as to avoid condensation.
repeat for remaining plates.

Seed Placement

draw horizontal line across plate with a center point roughly one inch from the perimeter of plate
place reference grid (link to pic or file) under plate, matching horizontal lines.
set pipeter to small volume(?) so that when fully released the seed mixture level remains within the lower angled section of the blue tip.
after pulling a small amount of seed (#3) into blue tip, remove tip.
remove lid off first plate and keeping the lid as a shield between you and the open plate.
to place seed, reference the grid below the agar and gently touch surface of agar with tip, so that only one seed is placed onto the agar. (Too little seeds in tip will result in no seeds coming out of tip, and too much seed mixture will cause a group of seeds to come out of tip onto agar). If seeds do not come easily from tip, pressing one's finger into top of tube may work to create pressure and dislodge seed.
place seeds at desired density.
replace lid and move to next plate. Repeat steps.
when all plates are finished, seal with parafilm tape.
plates can be placed horizontal or vertical in growth room

Keep head out of hood. Contamination from hair, mouth, nose, hands, etc. will infect culture plate with mold spores.

@@ Line 24: / Line 24: @@
 #once all are in hood, remove aluminum foil wrap from agar flask.
 #remove lid off first plate and keeping the lid as a shield between you and the open plate, pour agar into plate. Fill to halfway point, carefully avoiding air bubbles in media. Place lid right-side-up on hood surface, then set poured plate on top. Allow to cool before replacing lid as to avoid condensation.
-#Repeat for remaining plates.
+#repeat for remaining plates.
 Seed Placement