eamag/protocols-with-steps huggingface dataset:
TITLE: WIPI2d Expression and purification
AUTHORS: lmstrong 

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">WIPI2d expression and purification from HEK cells </div></div>

[STEPS]
?. [Expression]
Transfect   of HEK GNTI cells
300 mL
?. [Purification]
Resuspended pellet in 25mL of lysis buffer (50mM Hepes pH 7.45, 300mM NaCl, 1mM TCEP, 1% Triton X, PI cocktail)
?. [Expression]
Dilute PEI with Warm Hybridoma-SFM(1X)
?. [Purification]
Let rock at for
4 °C
?. [Purification]
Clarify lysate for
Centrifuge: 18000 33, 30 min, 4 10
?. [Purification]
Rock supernatant with equilibrated strep resin for  at
4 °C
?. [Purification]
Let supernatant FT gravity column
?. [Purification]
Wash with 50CV wash buffer (50mM Hepes pH 7.45, 300mM NaCl, 1mM TCEP)
?. [Purification]
Concentrate elution and inject onto pre-equilibrated S200 10/30 column (25mM Hepes pH 7.45, 150mM NaCl, 1mM TCEP)
?. [Purification]
Elute 2X with 5mL of wash buffer spiked with 5mM des-thiobiotin
?. [Purification]
Pool peak fractions, concentrate, snap freeze, and store at -80C
?. [Expression]
In a separate tube, dilute DNA with Hybridoma-SFM(1X)
?. [Expression]
Add PEI to DNA dilution. Incubate mixture for   at
37 °C
?. [Expression]
Add mixture to cells. Let cells grow for
?. [Expression]
Harvest Cells
Centrifuge: 2500 33, 20 min, 4 10
?. [Expression]
Wash pellet with cold PBS. Store pellet until Puification.
TITLE: K1 keto - (Top Weight Loss Supplement) Quickly Work !!*SPECIAL OFFER*
AUTHORS: Advanced  Appetite

[DESCRIPTION]
K1 Keto Life Reviews- 100% Legit Weight Loss Supplement With Quick Results! Get you into ketosis in minutes without the need to follow a severe eating routine. 
(Buy Now)=>> https://reviewbylogix.online/k5an

[STEPS]
TITLE: RNA extraction_Trizol method_Protocol
AUTHORS: Rui Zhang, Clara Huesing, Heike Muenzberg

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">The study goal is to identify the gene expression profile of interscapular brown fat (iBAT)-related ganglia (SG/T1 & T3) and inguinal white fat (Iwat)-related ganglia (T13/L1 & L2)</div></div>

[STEPS]
?. Take out the samples from the -80 °C freezer and keep them stay in the ice.
?. Add 500 µl Trizol (Ambion® by Life Technologies, Catalog number: 15596-026) to each sample (when adding the Trizol, add it toward the tissue so as to make sure the tissue is in the Trizol solution, instead of sticking on the tube well).
?. Use a hand-held homogenizer (PELLET PESTLE® Cordless Motor, motor cordless, kon 749540-0000, kimble pellet pestle motor, kimble chase company) with the sterilized (by autoclaving) pellet pestles (part number: 749520-0500, kimble chase company) to homogenize the samples in the fume hood (Use the pellet to locate the tissue in between the micro-centrifuge tube well and the tip and squeeze it, then turn on the homogenizer in the tube).
?. Add 1.7 µL Glycoblue (GlycoBlueTM, Ambion® by Life Technologies) Note: 300-fold dilution; the total volume of the solution for each sample is 500 µL, so after diluting 300-fold dilution, the needed volume of glycoblue is 500/300=1.7 µL to each sample.
?. Note: Change the homogenizer tip for each sample. After it’s done, check the solution again to make sure the tissue is completely homogenized.
?. Centrifuge 30 phase lock gel (PLG) at the maximum speed of the centrifuge machine for 1 minute.
?. Transfer homogenized samples to the PLG micro-centrifuge tubes and incubate the homogenized sample at room temperature for 5 minutes.
?. Add 100 µl phase separation reagent (BCP, molecular biology grade 1-bromo-3-chloropropane) (catalog number: BP 151), Shake vigorously by hand for 15 seconds (by putting another micro-centrifuge tube rack on top of the micro-centrifuge tube rack that contains these sample tubes, and then shake them). Incubate at room temperature for 2-3 minutes.
?. Then centrifuge the sample at 12,000×g for 15 minutes at 4 °C.
?. Transfer the upper aqueous phase into new RNase-free polypropylene micro-centrifuge tubes.
?. Add 0.25 ml 100% isopropanol to the aqueous phase, mix well and incubate at room temperature for 10 minutes.
?. Centrifuge at 12,000 ×g at 4 °C for 15 min.
?. Remove the supernatant and wash the pellet with 500 µl 75% RNase-free ethanol.
?. Vortex the sample briefly and centrifuge at 7500 ×g at at 4 °C for 5 minutes.
?. Discard the ethanol and dry the RNA pellet in the heat block at 55-60 °C (avoid the RNA from being over dried).
?. Resuspend the RNA pellet in 20 µl RNase free water and incubate in a heat block set at 55-60 °C for 10-55 minutes.
?. Measure RNA concentrations with NanoDrop and proceed to the downstream application, or store at -80°C.
TITLE: Cold Spicy Sichuan Noodles
AUTHORS: Hao Ye

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Inspired by Ba Ren (RIP), sour, a little sweet, numbing spiciness. I added in some lightly-pickled cucumbers for extra crunch.</div></div>

[STEPS]
?. [Pickled Cucumbers]
Prepare pickled cucumbers following linked recipe.Brine ingredients:  AB11 cuprice vinegar (unseasoned)21 cupwater33/4 cupsugar41 tbspsaltOther ingredients:  AB16persian cucumbers (julienned)Notes:We are going to combine these cucumbers with the noodles, so keep that in mind when determining how large the pieces should be. I like to slice the cucumber pieces to 2-2.5 inches long and so that each piece has a bit of peel attached.The brine is sufficient for 6 cucumbers, though you only need 3-4 for each 12 oz of noodles, so the rest can be reserved for later consumption. :) http://www.seriouseats.com/recipes/2013/10/quick-pickled-cucumbers-rice-vinegar-recipe.html
AB11 cuprice vinegar (unseasoned)21 cupwater33/4 cupsugar41 tbspsalt
AB16persian cucumbers (julienned)
These can be prepared ahead of time (i.e. the night before).
?. [Noodle Preparation]
Cook noodles following package instructions.Ingredients:  AB1~ 12 oz.noodles22 tbspsesame seed oil32 bunchesgreen onions (thinly sliced)
AB1~ 12 oz.noodles22 tbspsesame seed oil32 bunchesgreen onions (thinly sliced)
Which noodles? I like the Sun noodles ramen kits - shoyu or tantanmen flavors have the same noodles with the size and consistency I like, but you can experiment here.
?. [Sauce Preparation]
Mix ingredients together:  AB11/2 cupzhenjiang rice vinegar (Gold Plum 'chinkiang')25 tspplain white sugar
31 tspsalt41 tspsichuan peppercorn oil52 tbspsichuan pickled peppers64 clovesminced garlicvinegar:peppercorn oil:pickled peppers:
AB11/2 cupzhenjiang rice vinegar (Gold Plum 'chinkiang')25 tspplain white sugar
31 tspsalt41 tspsichuan peppercorn oil52 tbspsichuan pickled peppers64 clovesminced garlic
The seasoning can be varied to taste, but note that oil separation will occur; so it is necessary to be sure that the sauce is mixed well before tasting.
?. [Serving]
Mix sauce and noodles well and serve.
Ideally, we want all the components to be adequately chilled - normal fridge temperature is probably a bit too cold, but if serving to guests, maybe let everything rest at room temperature for ~5 minutes before mixing.
Separately, noodles and sauce will keep for about a day in the fridge (longer might be ok, but I'm not sure about food safety), so everything could be prepared the night before and mixed right before serving.
?. [Noodle Preparation]
After cooking, rinse noodles with cold water to cool it down.
?. [Noodle Preparation]
Drain, then mix well with the pickled cucumbers and sesame seed oil, which will help prevent sticking while cooling in fridge.
?. [Noodle Preparation]
Top with thinly sliced green onions. Cover and chill in fridge while preparing sauce.
TITLE: Preparation of BG-11 media Agar 
AUTHORS: Ashwinuday  

[DESCRIPTION]
This is for making solid media of BG-11

[STEPS]
1. To make BG-11 Agar plates, add Agar-Agar Type-1 equivalent to 1.5% (w/v) of liquid media and autoclave.
2. Then before it solidifies (when the flask is bearable to touch), add the required antibiotics to the media.
3. Pour ~30-40 ml into plates soon after.
4. Keep the plates opened in the hood.
5. Once it solidifies in the plates then close those and keep the plates inverted into a cold storage.
TITLE: Chitin binding + Bradford assay
AUTHORS: Robert Hooftman

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">This protocol can be used to measure the binding capacity of chitin binding proteins and visualizing this using a Bradford assay. The protocol has been adapted from:</div><div class = "text-block">F. Labroussaa, A. R. Zeilinger, and R. P. P. Almeida, “Blocking the Transmission of a Noncirculative Vector-Borne Plant Pathogenic Bacterium,” Mol. Plant-Microbe Interact., vol. 29, no. 7, pp. 535–544, Jul. 2016.</div></div>

[STEPS]
?. [Chitin binding assay]
First, prepare the following buffer:Chitin buffer  ABC1KH2PO4&nbsp;2 mM&nbsp;(0,272 g/L)&nbsp;2Na2HPO4&nbsp;8 mM&nbsp;&nbsp;&nbsp;(1,424 g/L in case of Na2HPO4&nbsp;* 2 H2O)&nbsp;3KCl&nbsp;2 mM&nbsp;&nbsp;(0,149 g/L)&nbsp; Preparation of chitin solution. Make a chitin stock of 10 mg/ml in the chitin buffer.
ABC1KH2PO4&nbsp;2 mM&nbsp;(0,272 g/L)&nbsp;2Na2HPO4&nbsp;8 mM&nbsp;&nbsp;&nbsp;(1,424 g/L in case of Na2HPO4&nbsp;* 2 H2O)&nbsp;3KCl&nbsp;2 mM&nbsp;&nbsp;(0,149 g/L)&nbsp;
?. [Chitin binding assay]
In a 2 ml microfuge tube, add 50 ug of proteinin solution (chitin binding protein or a control, such as BSA).
?. [Chitin binding assay]
Add500 ug of chitin from the chitin solution (50 ul in case of a 10 mg/ml stock solution). Make sure the solution is being stirred while pipetting to make sure the solution is completely homogenized!
?. [Chitin binding assay]
Adjust total volume to 500 ul.
?. [Chitin binding assay]
Incubate themicrofuge tubeat room temperaturefor 1 hour while shaking (600 rpm on Eppendorf shaking block) to prevent chitin from precipitating.
?. [Chitin binding assay]
After incubating, centrifuge for 3 minutes in a tabletop centrifuge at 13000 x g.
?. [Bradford assay]
Make protein standards, with a known concentration (0 ug/ml, 1.25 ug/ml, 2.5 ug/ml, 5 ug/ml, 10 ug/ml, 15 ug/ml and 20 ug/ml).
?. [Bradford assay]
Make dilutions for your sample (e.g. 5x and 20x).
?. [Bradford assay]
Add Bradford Reagent (Sigma) 1:1 to your samples.
?. [Bradford assay]
Let the samples incubate for 10 minutes at room temperature.
?. [Bradford assay]
Transfer the solutions + reagent to 1 ml cuvettes.
?. [Bradford assay]
First, measure the A595 (absorabance at 595 nm) for the sample containing 0 ug/ml protein. Use a photospectrometer. This will be the blank.
?. [Bradford assay]
Measure the A595 for the rest of the protein standards
?. [Bradford assay]
Use these to make a graph of A595 (y-axis) vs protein concentration (x-axis) and make a trendline + formula.
?. [Bradford assay]
Measure the A595 of your protein samples. Make sure that the absorbance is within the value range of the standards. If not, measure the diluted sample(s) instead.
?. [Bradford assay]
Calculate the protein concentration of your sample with the formula obtained from the trendline. Make sure to adjust your concentration in case you measured a diluted sample!
?. [Bradford assay]
Plot your data
TITLE: Latex beads migration assay test
AUTHORS: Jorge Fernández

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">The following protocol details how to test the migration of conjugated latex beads trough different nitrocellulose membranes.</div></div>

[STEPS]
?. [Membrane Preparation]
Prepare two FF170HP strips  and three FF80HP strips of 1cm wide x 4 cm long.Wax print the microfluidic membranes following the protocol mentioned in the guideliness section.
?. [Membrane Preparation]
Block one FF170HP and two FF80HP membranes by immersion in 0.2 % BSA solution in PBS. Let the membranes dry for 1h at room temperature, and let them on a dissecator at 4ºC overnight.
?. [ Migration test]
Aspire  20 µL of  conjugated latex beads stock, and pipette them on the sample deposition area of the sealed membranes. It's recommendable placing the strips with a briefly inclination degree, avoiding the sample to fall down trough the membrane surface. (Placing the sample deposition region on the down region).
?. [ Migration test]
Wait 15 minutes until the liquid in the sample has migrated completely. Results can be directly visualized. Wait until the membranes have dried to manipulate them.
TITLE: Build your own calculus
AUTHORS: Bjørn Peare  Bartholdy, a.g.henry

[DESCRIPTION]
This collection contains protocols for growing a calcifying oral biofilm model. Protocols include artificial saliva, a mineralising solution (CPMU), an alpha-amylase activity assay, and a biofilm growth protocol with starch treatments.

The purpose of these protocols is to explore fundamental aspects of using dental calculus to infer dietary patterns in past populations. The main protocol contains starch treatments to look at the incorporation and extraction of starch granules from dental calculus, but can also be used to look at other dietary components, such as various proteins, other plant extracts, and whatever else may have been consumed in the past.

Protocols are modified protocols from Sissons et al. (1991) and Extercate et al. (2010)

[GUIDELINES]
The protocols should be conducted in a sterile environment to avoid contamination of the biofilm with external contaminants. 

If starch is used for treatments, it is important to work in a starch-free lab and include control samples in each plate to test for external contamination and cross-contamination between wells.

[STEPS]
TITLE: El-cheep-o miniprep
AUTHORS: Magdalena Julkowska

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">This is a protocol for cheep miniprep isolation. Be aware that the DNA quality might not be sufficient to use this protocol for cloning purposes. It works very well for the digestion analysis though. </div></div>

[STEPS]
?. ·Spin 3ml of the overnight culture to pellet cells (1.5-ml for time) @13000 rpm for 5 min
?. ·Remove the supernatant
?. ·Add 250 ml of sol P1+Rnase 5ml and vortex to resuspend cells
?. ·Take 15 min @ room temperature
?. ·Add 250 ml of sol P2 and mix gently
?. ·Take 5 min @ room temperature
?. ·Add 350 ml of sol P3 and mix gently
?. ·Leave 10 min on ice
?. ·Spin @ the max speed for 10 min
?. ·Take 750 ml of the supernatant and put in a new eppendorf tube and add in this tube 750 ml of isopropanol
?. ·Mix well
?. ·Spin for 30 min @ 14000 rpm
?. ·Remove the supernatant
?. ·Spin again for other 10 min
?. ·Take @ 37°C for 10 min
?. ·Add 50 ml of H2O
?. ·Store @ -20°C
TITLE: Salmonella purification
AUTHORS: Angel  Justiz-Vaillant, Suzette E Curtello

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><span>The United States veterinarian Daniel E Salmon discovered the genus </span><span style = "font-style:italic;">Salmonella</span><span>. </span><span style = "font-style:italic;">Salmonella </span><span>is non-spore forming and it may survive without oxygen. This bacterial strain is mainly associated with animals. </span><span style = "font-style:italic;">Salmonella </span><span>is ubiquitous and may be asymptomatic which has generated great health concern.This bacterium is associated with life threatening diseases among human and animals.Gastroenteritis is one such disease which may resolve itself within five days in healthy individuals.Immuno compromised individuals and the young are at great risk as this may progress to secondary systemic complication.</span></div><div class = "text-block"><span style = "font-style:italic;">Salmonella</span><span> serotype exceeds two thousand which may be differentiated from each other by the International Kauffmann-white Scheme Serotyping Manual.Surface antigens produced by salmonellae are used in the determination of isolated strain.Determination of the “O” antigen group associated with the carbohydrate constituent of the lipopolysaccharide membrane of the bacterium precede the identification of the protein “H” antigen which is linked to the tail like flagella of the bacterium.The two existing phases of </span><span style = "font-style:italic;">Salmonella</span><span>may be referred to as specific and nonspecific which function as motile and non motile phases.The existing phase of </span><span style = "font-style:italic;">Salmonella</span><span>will determine the “H” antigens that are produced.An additional “Vi” antigen is associated with virulent encapsulated strains of </span><span style = "font-style:italic;">Salmonella</span><span> Typhi.</span></div></div>

[STEPS]
?. A 1:9 Salmonella suspension was made in buffered peptone water and incubated overnight at 37ºC.
?. One ml of pre-enrichment broth was transferred to a 1.5 ml micro-centrifuge tube and centrifuged for ten minutes at 14,000 x g (Eppendorf Model 5424).
?. The supernatant was carefully discarded.
?. The pellets were re-suspended in 300 µl sterile PCR grade water by vortexing.
?. The tube was again centrifuged at 14,000 x g for five minutes.
?. The supernatant was discarded with care.
?. The pellets were again resuspended in 300 µl PCR grade water by vortexing.
?. The microcentrifuge tube was incubated at 100ºC for 15 minutes and immediately chilled on ice.
?. The tube was centrifuged at 14,000 x g at 4ºC.
?. The supernatant was transferred to a new tube and incubated at 10 min at 100ºC  then chilled immediately on ice.
?. The supernatant was stored at -20 ºC.
TITLE: Antigen Presentation Protocol
AUTHORS: Lilia Rodriguez

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">This protocol details methods for 3-day Antigen Presentation Assay.</div></div>

[STEPS]
?. [Day 1]
Prepare a 96 well plate:Label appropriately (experimental condition – LPS/IFNg controls, infection controls, etc. ) Duplicate labelled wells for the peptide control
?. [Day 1]
Scrape RAW cells and resuspend (pipette up and down 8x).
?. [Day 1]
Count RAW cells and adjust concentration to 0.75 million cells/ml.
?. [Day 1]
Add  to each well.
[RAW cells]
?. [Day 1]
Add LPS/IFNg accordingly.
?. [Day 1]
Incubate for  at .
[with 5% CO2]
?. [Day 2]
Prior to beginning, check the cell’s media (red/orange = good, yellow = bad).
?. [Day 2]
Prepare  at  ().
[PFA in PBS]
37 Room temperature
?. [Day 2]
Discard supernatant in the 96 well plate and add .
[1% PFA]
?. [Day 2]
Incubate at  for .
37 Room temperature
?. [Day 2]
Prepare 2E2 cells while RAW cells are in 1% PFA:
?. [Day 2]
Pour flask into 50 ml conical and centrifuge at .
Centrifuge: 1500 33, 3 min
To keep more 2E2 cells growing, add   back into their flask and incubate at  .
?. [Day 2]
Count 2E2 cells and adjust concentration to 0.4 million cells/ml.
?. [Day 2]
Add  to each well in the 96 well plate and discard immediately.
[wash buffer]
Wash buffer = DMEM with 0.1M glycine, + 10% iFBS.
?. [Day 2]
Repeat 2x more: Add  to each well in the 96 well plate and discard immediately. (Wash 1/2)
[wash buffer]
?. [Day 2]
Repeat 1x more: Add  to each well in the 96 well plate and discard immediately. (Wash 2/2)
[wash buffer]
?. [Day 2]
Add  per well in ½ of the samples (the non-peptide ones). Add an extra row of 2E2 cells alone for a negative control.
[2E2 cells]
?. [Day 2]
Dilute stock peptide (1 µg/ml) 5000 fold in the 2E2 cells (final concentration: ).
?. [Day 2]
Add  to the remaining wells.
[peptide + 2E2 cell solution]
?. [Day 2]
Incubate at  for .
[, 5% CO2]
[(NO LONGER)]
?. [Day 3]
Prepare 1x lysis buffer (stock = 5x, with triton, ) in dH2O.
?. [Day 3]
Add  in .
[1M stock of DTT]
[lysis buffer]
?. [Day 3]
Prepare CPRG buffer ().
?. [Day 3]
Centrifuge 96 well plate at .
Centrifuge: 2200 33, 1 min
?. [Day 3]
Add  to all wells.
[lysis buffer]
?. [Day 3]
Incubate at  for  (up to 20 minutes MAX).
37 Room temperature
?. [Day 3]
Prepare CPRG (recipe below = for each well)
[CPRG buffer]
[water]
[CPRG]
?. [Day 3]
When lysis is done, add  / well.
[CPRG solution]
?. [Day 3]
Incubate either at  or 37°C (37 speeds up reaction to about ~ 20 minutes for peptide samples).
37 Room temperature
?. [Day 3]
Transfer  to a new plate.
[colored solution]
Take care not to transfer debris or make bubbles.
?. [Day 3]
Take reading at 595 nm or 570 nm.
* To stop reaction to leave overnight, incubate at  .
TITLE: General Fungal DNA Extraction
AUTHORS: Angie Macias, Matthew T Kasson, Brian Lovett

[DESCRIPTION]
This is a routine protocol for extracting DNA from various fungi. This extraction method is suitable for follow-up molecular work such as PCR amplification.

[STEPS]
SECTION: Before you begin
1. Turn on hot water bath, set to 65 °C.

SECTION: Before you begin
2. Pull two Eppendorf 1.5 mL centrifuge tubes per sample.

SECTION: Before you begin
2.1. Label both sets of tubes with (short) sample names.
SECTION: Before you begin
2.2. Label one tube set for each sample with an "I" for  .
  

SECTION: Before you begin
3. Add 200 µL of   (or () to tubes without "I".

SECTION: Before you begin
4. Add 600 µL of   to tubes labeled with "I".

SECTION: Before you begin
5. Place tube with   into 65 °C water bath.

SECTION: Extraction Protocol
6. Sterilize some metal scrapers with flame and 95 % (v/v)   .

SECTION: Extraction Protocol
7. Add 1/2 pea-sized amount of fungal tissue (young hyphae) to each tube containing  .

SECTION: Extraction Protocol
7.1. Flame-sterilize and cool scrapers between samples.
SECTION: Extraction Protocol
8. Macerate each sample with a new, sterile micropestle until tissue is homogenous.
SECTION: Extraction Protocol
10. Add tubes to a floating rack to allow samples to incubate directly in 65 °C water bath for 30 min.

SECTION: Extraction Protocol
11. Remove samples and vortex for 3 s before returning to 65 °C water bath for 30 min.

SECTION: Extraction Protocol
12. Remove samples and allow them to cool on the bench for 5 min.

SECTION: Extraction Protocol
13. Add 200 µL of   to each tube and vortex for 10 seconds.

SECTION: Extraction Protocol
14. Centrifuge samples for 3 min at 14.000 rpm.

 

SECTION: Extraction Protocol
15. Using a P1000 micropipette, transfer supernatant to each tube containing   and gently mix by inversion.
 


SECTION: Extraction Protocol
16. Centrifuge for 1 min at 14.000 rpm.

SECTION: Extraction Protocol
17. Carefully pour off the supernatant into waste container.
 

SECTION: Extraction Protocol
18. Add 600 µL of 70 % (v/v)   to each tube and mix gently by inversion.

SECTION: Extraction Protocol
19. Centrifuge for 1 min at 14.000 rpm.

SECTION: Extraction Protocol
20. Repeat Step 16.
SECTION: Extraction Protocol
21. Open and invert tubes onto a clean paper towel.
 

SECTION: Extraction Protocol
22. Add 100 µL of warmed   to each tube.

SECTION: Extraction Protocol
23. Store fully-labeled tubes in a box (not a tube rack) in the -20 °C freezer.

SECTION: Extraction Protocol
11.1. Place a sufficient aliquot of   in water bath to warm for Step 21.

SECTION: Extraction Protocol
9. Add 400 µL of   (to 600 µL total volume added).
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p>In order to take chlorophyll fluroscence images of cowpea accessions, leaf disks need to be collected and placed randomly onto a leaf disk tray. This protocol explains how.</p>

[STEPS]
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?.