Marine Biology

Marine Biology kdorfman Mon, 01/15/2024 - 17:32

Bio 424 Spring 2024 (Akiko Okusu)

Materials Request

Prep Instructions

Growth chamber spawning program

Growth chamber spawning program kdorfman Fri, 05/17/2024 - 19:17

Basic Parameters:

Light from 9 pm Sunday to 1 pm Monday, otherwise dark

Warm from 9 pm Sunday til 9 am Monday, otherwise cool

Day time temp light level
M 13:00 18 0
Sun 21:00 25 3
M 9:00 18 3
M 13:00 18 0

Schedule 3 programs, like this:

Program 1, repeat 6 times:

Day time temp light level
M 00:00 18 0

Program 2 (spawning), do once, following 6x Program 1

Day time temp light level
Sun 00:00 18 0
21:00 251 3

Program 3, do once

Day time temp light level
M 00:00 25 3
9:00 18 3
13:00 18 0

Return to Program 1

  1. There is a temperature spike when the light comes on, so leave the temp setting at 18 for an hour (until 22:00), then raise it to 25. ↩︎

Sea Water for 424

Sea Water for 424 kdorfman Mon, 01/15/2024 - 18:14

2 carboys SeaWater (31-35ppt salinity):

  • 31-35g InstantOcean Reef Salt in 1 L
  • Submerge a bubbler or a pump so that it is constantly being mixed

Nematostella SW is 1/3 ASW

F/2 Medium for algae (424)

F/2 Medium for algae (424) kdorfman Mon, 01/15/2024 - 18:16

5 liters of f/2 medium for algae.

L1 Medium Kit from Bigelow NCMA contains the nitrate, phosphate, trace minerals, and vitamins. Add the antibiotics from prep room freezer stocks.

Re-inoculate once during week of 1/15

TA & Students will re-inoculate once monthly

50 mL/month x 4 months x 16 students = 3200 mL

Make 4 L for the semester.

To make (in a sterile hood)L F/2 medium for algae
add: mL NaNO3 (75g/L)
add: mL NaHPO4 (5 g/L)
add: mL trace minerals
add: mL vitamins
add: mL kanamycin
add: mL ampicillin
add: mL streptomycin

Plus ASW to final volume

Filter sterilize

F/2 medium for algae

Trace minerals for F/2

Trace minerals for F/2 kdorfman Mon, 06/24/2024 - 15:46

Buy L1 Medium Kit from Bigelow NCMA

Replacement trace minerals solution

Component Concentration in final medium
Na2EDTA · 2H2O 1.17 x 10-5 M
FeCl3 · 6H2O 1.17 x 10-5 M
MnCl2·4 H2O 9.00 x 10-7 M
ZnSO4 · 7H2O 8.00 x 10-8 M
CoCl2 · 6H2O 5.00 x 10-8 M
CuSO4 · 5H2O 1.00 x 10-8 M
Na2MoO4 · 2H2O 8.22 x 10-8 M
H2SeO3 1.00 x 10-8 M
NiSO4 1.00 x 10-8 M
Na3VO4 1.00 x 10-8 M
K2CrO4 1.00 x 10-8 M

Vitamins for F/2

Vitamins for F/2 kdorfman Mon, 06/24/2024 - 15:51

Buy L1 Medium Kit from Bigelow NCMA

Replacement Vitamin Stock Solution

Component Concentration in Final Medium
thiamine · HCl (vit. B1) 2.96 x 10-7 M
biotin (vit. H) 2.05 x 10-9 M
cyanocobalamin (vit. B12) 3.69 x 10-10 M