Planting guide
Planting guide kdorfman Mon, 01/16/2012 - 15:48Check seed stocks before planting - some germinate better than others!!
Hydroponics video: https://www.youtube.com/watch?v=c9neVLaS63c
Hydroponics paper: http://www.plantmethods.com/content/9/1/4
2012 Schedule
2012 Schedule kdorfman Fri, 12/16/2011 - 17:10Potting on soil
date | wks before | Lab | purpose | per pot | pots/pair | # pots |
---|---|---|---|---|---|---|
1/4 | 8 | 0.3 | demo | 3-5 | 1 | 10 |
1/11 | 7 | 0.3 | demo | 3-5 | 1 | 10 |
3 | 1.1 | DNA | lots! | 4 | 40 | |
1/18 | 6 | 0.3 | demo | 3-5 | 1 | 10 |
2 | 1.1 | DNA | lots! | 4 | 40 | |
1/25 | 5 | 0.3 | demo | 3-5 | 1 | 10 |
2/1 | 4 | 0.3 | demo | 3-5 | 1 | 10 |
2/8 | 3 | 0.3 | demo | 3-5 | 1 | 10 |
2/15 | 2 | 0.3 | demo | 3-5 | 1 | 10 |
2/27 | 6 | 5.3 | projects | 3-5 | 1 | 10 |
3/6 | 5 | 5.3 | projects | 3-5 | 1 | 10 |
3/13 | 4 | 5.3 | projects | 3-5 | 1 | 10 |
3/20 | 3 | 5.3 | projects | 3-5 | 1 | 10 |
3/27 | 2 | 5.3 | projects | 3-5 | 1 | 10 |
Planting on plates
date | for Lab | purpose | # plates | notes |
---|---|---|---|---|
2/22 | 0.3 | demo | 10 | 1 weeks before lab |
3/26 | 5.3 | projects | 10 | 2 weeks before lab |
4/3 | 5.3 | projects | 10 | 1 weeks before lab |
Calendar Spring 2012
Calendar Spring 2012 kdorfman Tue, 12/20/2011 - 16:29January 2012
Sun | Mon | Tue | Wed | Thurs | Fri | Sat |
---|---|---|---|---|---|---|
1 Holiday | 2 | 3 | 4 Plant | 5 | 6 | 7 |
8 | 9 | 10 | 11 Plant | 12 | 13 | 14 |
15 | 16 Holiday | 17 | 18 Plant | 19 | 20 | 21 |
22 | 23 | 24 | 25 Plant | 26 | 27 | 28 |
29 | 30 | 31 |
February 2012
Sun | Mon | Tue | Wed | Thurs | Fri | Sat |
---|---|---|---|---|---|---|
1 Plant | 2 | 3 | 4 | |||
5 | 6 | 7 | 8 Plant | 9 | 10 | 11 |
12 | 13 | 14 | 15 Plant | 16 | 17 | 18 |
19 | 20 Holiday | 21 | 22 | 23 | 24 | 25 |
26 | 27 Plant | 28 | 29 |
March 2012
Sun | Mon | Tue | Wed | Thurs | Fri | Sat |
---|---|---|---|---|---|---|
1 | 2 | 3 | ||||
4 | 5 | 6 Plant | 7 | 8 | 9 | 10 |
11 | 12 | 13 Plant | 14 | 15 | 16 | 17 Spring Break |
18 | 19 | 20 Plant | 21 | 22 | 23 | 24 |
25 | 26 | 27 Plant | 28 | 29 | 30 | 31 |
April 2012
Sun | Mon | Tue | Wed | Thurs | Fri | Sat |
---|---|---|---|---|---|---|
1 | 2 | 3 | 4 | 5 | 6 | 7 |
8 | 9 | 10 | 11 | 12 | 13 | 14 |
15 | 16 Holiday | 17 (Mon) | 18 | 19 | 20 | 21 |
22 | 23 | 24 | 25 | 26 | 27 | 28 |
29 | 30 |
May 2012
Sun | Mon | Tue | Wed | Thurs | Fri | Sat |
---|---|---|---|---|---|---|
1 Last day | 2 | 3 | 4 | 5 | ||
6 | 7 | 8 | 9 | 10 | 11 | 12 |
13 | 14 | 15 Grades due | 16 | 17 | 18 | 19 |
20 | 21 | 22 | 23 | 24 | 25 | 26 |
27 | 28 Holiday | 29 | 30 | 31 |
On plates
On plates kdorfman Mon, 01/16/2012 - 15:45Planting on Plates
Make MS plates
- To get weakling mutants started before potting in soil
- Or to plant seeds on specified media
- Or to see roots
Seed Preparation
Seeds must be sterilized before they are put on sterile plates.
After sterilization,
- Make a line of seeds in agarose above the midline of the plate, either
- either on the agar directly
- on flattened cellophane
- Tape plates shut with Micropore surgical tape
- Stack plates flat
- Refrigerate the plates for 2 days in the dark
- Move plates to growth chamber
- Stand plates on edge under the lights. Tape several together.
0.1% agarose for plating seeds
- 100 mL sterile ddH2O
- 0.1 g agarose . autoclave to melt; shake bottle after autoclaving
Sterilize seeds with EtOH & Tritonx
Sterilize seeds with EtOH & Tritonx kdorfman Mon, 02/14/2022 - 18:31sterilize seeds in a solution of
- 0.5 mL 70% EtOH,
- 0.05% TritonX-100,
- ~3 min mild agitation.
Let seeds settle, remove liquid
Replace with sterile water, 3X
Replace water with sterile 0.1% agar, 3X
Plate in a line, either
- directly on agar, one seed per tiny drop or
- on flattened cellophane (wet it if necessary)
Wrap plates with micropore tape
stack plates flat
cover in foil, refrigerate, 48 hours
Sterilize seeds with EtOH then dry
Sterilize seeds with EtOH then dry kdorfman Mon, 02/14/2022 - 18:39Put appropriate number of seeds in a sterile microfuge tube
Sterilize filter paper with EtOH, let dry in sterile hood (can set it on top of an open sterile petri plate cover
Add 95% EtOH to seeds for 5 min, dump seeds onto filter paper
let dry, tap onto agar of an MS plate
Sterilize seeds with bleach
Sterilize seeds with bleach kdorfman Mon, 02/14/2022 - 18:37- Add ~1mL of bleach solution to each tube
- Cap and shake
- Leave tube on its side for 20-30 min, shaking periodically
- Put upright after last shake (so seeds settle)
- In laminar flow hood
- Use a pipetman that has been sprayed with ethanol and air-dried in the hood
- Remove as much bleach solution as possible from the tube
- Add ~1 mL sterile ddH2O, mix and let seeds settle again
- Do at least 4 washes, to dilute out the bleach
- Add ~1mL sterile 0.1% agarose (so seeds will be suspended in the liquid)
Bleach solution for sterilizing seeds
- 7 mL sterile ddH2O (must be freshly made!)
- 3 mL Chlorox
- 1 uL Triton X-100 (detergent to help get bleach into seed crevices)
0.1% agarose for plating seeds
- 100 mL sterile ddH2O
- 0.1 g agarose . autoclave to melt; shake bottle after autoclaving
On soil
On soil kdorfman Mon, 01/16/2012 - 15:43Arabidopsis planting guide - Plants on soil
Routine weekly planting to produce plant material for observation or DNA extraction
- Pots
- 6 pots in a big flat with no holes
- fill with clump-free potting soil, leaving some space at the rim
- Treat with Gnatrol for fungus gnats
- saturate with hot water the night before Teddi comes
- Arrange for Teddi to do Gnatrol treatment
- OR Heat in oven
- pre-heat oven to 225F (=~107C)
- Put soil-filled pots in aluminum lasagne pan
- Cover with foil
- insert thermometer in one pot near center
- Heat to 180F (=~82C)
- leave in oven for 30 min.
- leave covered until ready to plant
- Seeds for DNA
- Start a set 4 weeks before needed, another set 3 weeks before
- Columbia wild type (col-0)
- Scatter seeds on surface of soil (remoisten before if it is dry)
- Put on plastic cover
- Put in refrigerator, keep dark (this synchronizes germination)
- Transfer to growth chamber on the third day
- If no room in fridge, put seeds in microfuge tube, add water, refrigerate in dark 2 days, then plant
- Seeds for demo or experimental plants
- Put seeds in microfuge tube, water, refrigerate, keep dark
- 3rd day: pour into small beaker, add more water
- Deposit one seed to each corner of the pot, plus one in the middle with a transfer pipet.
- Transfer to growth chamber
- Growth Chamber
- 22C, 16H light, schedule continuous program
- Water bottom flat ~3x per week
- Remove cover after plants are well established.
hydroponics
hydroponics kdorfman Thu, 05/22/2014 - 21:33http://www.plantmethods.com/content/9/1/4
http://www.bio-world.com/productinfo/3_44_292/124429/Magenta-GA-Plant-C…
2015 attempt
Make 1x MS, pH to 5.6
Split: most as growth medium, a little bit for 0.7% agar (err on the low end), sterilize
Cut lids off microfuge tubes, poke a hole in each, sterilize in a beaker
line lids up, flat side down, on a strip of scotch tape
fill with 0.7% agar.
put lids into floatie or other rack.
put 1 sterilized seed in each hole. (in 0.1% agar)
Wrap with saran wrap
Cover tightly with foil, stratify in refrigerator for 2-3 days
Remove foil, put container in growth chamber at 16h day, 22C
Transfer lid to 50 mL tube with hole drilled in it at ~3 weeks.
Aerate or stir medium