eamag/protocols-with-steps huggingface dataset:
TITLE: Broccoli and Chicken Stir Fry
AUTHORS: Andy Ouyang

[DESCRIPTION]
A simple broccoli and chicken stir fry to create as college students with limited cooking experience and ingredients

[STEPS]
SECTION: Prepping Stage
1. 
 
In this step, you will have to gather your ingredients:
2 heads of broccoli
3 defrosted chicken breasts
2/3 cup of soy sauce
1/3 cup of oyster sauce
honey (as much as desired)
2 tablespoons water
2 tablespoons cornstarch
2 teaspoons vegetable oil

4. Stir soy sauce and oyster sauce together in a bowl. Mix in as much honey as desired (all portion sizes for the sauce are modifiable to suit personal taste)


SECTION: Prepping Stage
2. Chop the heads of the broccoli into florets and wash thoroughly
SECTION: Prepping Stage
3. Cut the chicken breasts into 1-inch-sized cubes
5. Mix water and cornstarch together in a small bowl and stir until the cornstarch dissolves completely.
SECTION: Cooking Stage
6. Heat the vegetable oil in a large skillet over high heat for 1 min 
 


SECTION: Cooking Stage
7. Fry chicken in hot oil until chicken is no longer pink in the center, for 5 min 

SECTION: Cooking Stage
8. Stir in broccoli with chicken and sauté until broccoli is hot, for 5 min .

SECTION: Cooking Stage
7.1. If the chicken is not ready yet, fry it for another 2 min 

SECTION: Cooking Stage
9. Push the chicken and vegetable mixture to the side of the skillet. Pour the soy sauce mixture into the vacant part of the skillet. Stir cornstarch slurry into the soy sauce mixture until the color is consistent.
SECTION: Cooking Stage
10. Move chicken and vegetables back into the center of the pan; sauté until sauce thickens and coats chicken and vegetables, for 5 min .

SECTION: Cooking Stage
11. Move your broccoli and chicken stir-fry onto a plate and serve.
TITLE: DNA extraction - mouse tails - phenol-chloroform 
AUTHORS: Wayne Crismani

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">This protocol is taken from the URL indicated below from the Jackson Laboratories</div><div class = "text-block">This protocol yields a highly purified DNA preparation from mouse tail biopsies suitable for next-generation sequencing.</div></div>

[STEPS]
?. Remove 0.5 mm of tail into polypropylene microfuge tube (do not mince). (The tubes must have tight-fitting caps, so that there are no leaks in steps 3 and 7 below.)
?. Add 0.5 ml DNA digestion buffer with proteinase K added to 0.5 mg/ml final concentration. (0.5 mg/ml is a high concentration and can probably be reduced.) DNA digestion buffer: •	50 mM Tris-HCl pH 8.0 •	100 mM EDTA pH 8.0 •	100 mM NaCl •	1% SDS
?. Incubate overnight at 50-55 °C with gentle shaking. (At this step, mechanical agitation greatly aids complete disruption of the tail.)
?. Quick-spin tubes to get solution off inside of cap.
?. Fill inside well of microfuge tube cap with vacuum grease. (We use Dow Corning high-vacuum grease and a 10cc syringe to dispense.)
?. Add 0.7 ml neutralized phenol/chloroform/iso-amyl alcohol (25:24:1).
700 µl
?. Mix fairly vigorously at 4C. (Do NOT vortex; we use a clinical rotator for 1 hour.)
?. Spin in microfuge  at top speed 5 minutes and transfer 0.5 ml of the upper phase to new microfuge tube. (Use P1000 for transfer, and draw the aqueous phase gently through tip several times after transfer if the DNA is still in large, gelatinous mass.)
?. Add 1 ml 100% ethanol at room temperature and invert (using clinical rotator if you wish) until DNA precipitate forms. (approximately 1 minute).
1 mL
?. Spin in microfuge 5 minutes and carefully remove and discard supernatant.
?. Add 0.5-1 ml 70% ethanol (-20 °C) and invert several times.
?. Spin in microfuge 5 minutes and care fully remove and discard supernatant.
?. Quick-spin tubes and remove last drop of ethanol solution with 25 µl capillary tube.
?. Air dry at room temperature or in dessicator (overnight if you wish).
?. Add 100-200 µl TE buffer and incubate at 65 °C for 15 minutes to resuspend DNA. Draw DNA through P1000 tip after 65 °C incubation to aid in suspension if you wish.
?. Use 10-20 µl for restriction enzyme digest.
?. Total yield is approximately 20-50 µg DNA, 0.1-0.25 µg/µl.
TITLE: DNA extraction from insect gut-dwelling fungi
AUTHORS: Yan Wang

[DESCRIPTION]
This protocol is good for DNA extraction from microbial fungi isolated from aquatic insect guts. It works for small input tissues (starting from one fungal thallus). The product can be used for PCR and Sanger sequencing directly, which is ideal for fungal barcode analyses. This protocol is modified from a method described in Gottlieb and Lichtwardt 2001 (Mycologia Vol. 93, No. 1, pp. 66-81).

[STEPS]
1. Suspend fungal thalli and spores in a 1.5 mL centrifuge tube with 2x CTAB buffer (200 µL). 

2. Freeze and thaw the sample three times by submerging the tube into liquid nitrogen and incubating it at 65 °Cusing a heat block.

3. After the final thaw, crush the fungal tissues using a disposable pellet pestle (for 1.5 mL microtube).
4. Cool the sample for 1 min on ice.

5. Add 4 µL Proteinase K 20 mg/mL to the tube and mix them by inverting the tube three times.

6. Warm the tube at 65 °Cfor 30 min using a heat block.

7. Centrifuge at 12000 x g, 2 min and transfer the supernatant to a new tube.

8. Add Chloroform: Isoamyl alcohol (24:1) with an equal volume of the supernatant.
9. Shake the mixture slowly for 10 min on a rotator and leave the tube on the rack for 2 min at room temperature.

10. Centrifuge at 13000 x g, 4 min and transfer the top layer to another tube with wide-bore tips, avoiding breaking DNAs.

11. Transfer the supernatant to a new tube and repeat steps 8-10.
12. Add 4 µL RNase A (10 mg/mL) and incubate at 37 °C for 40 min.

13. Add Isopropanol (stored at -20 °C) with a 66% volume of the product from the last step and mix by inversion.

14. Leave the tube at 4 °C for at least 300 min (or overnight) until fully precipitated.

15. Centrifuge at 13000 x g, 10 min with the cap opening side pointing to the centrifuge center.

16. Immediately dump the supernatant and transfer the tube (with the cap open & upside down) on the paper towel to absorb the remaining supernatant.
17. Leave the cap open on rack.
18. Add 150 µL 70% ethanol (stored at -20 °C) to the tube.

19. Cap it and use the finger to tap the tube gently.
20. Centrifuge at 13000 x g, 1 min.

21. Repeat step 16.
22. Leave the tube at 37 °C with the cap open and a piece of Kimwipe paper on top for 30 min or until the DNA becomes dry.

23. Add 50 µL H2O to the dried DNA in the tube and wait for 10 min.

24. Pipette and store the DNA in the tube at -20 °C.
TITLE: No Title
AUTHORS: 

[BEFORE_START]
If you are collecting flies from a field site to begin your house fly colony, be sure to keep field-collected adult flies away from any existing fly colonies or colony supplies.  Field flies are likely to carry several fungal and bacterial pathogens that can infect your existing fly colonies.  Instead, hold field flies separately and collect eggs from these flies that can then be placed into your colony rearing rooms without infecting other colonies.

[GUIDELINES]
This protocol is designed for large scale laboratory rearing of house flies at very low cost.

[STEPS]
TITLE: Liposomes
AUTHORS: BOC      Sciences

[DESCRIPTION]
Liposomes are spherical vesicles with internal cavities and composed of one or more phospholipid bilayers. Their structural components are phospholipids or synthetic amphiphilic combined with cholesterol and other sterols, which can affect membrane permeability. Liposomes can encapsulate hydrophilic or lipophilic molecules. Their applications are now well established in various fields such as medicine, biomolecules, and gene delivery, and have been studied as DDS (drug delivery system) to enhance treatment safety and efficacy of the agent.

[STEPS]
1.
TITLE: Intracardiac perfusion with buffer for anatomical studies
AUTHORS: Janet R Keast, Peregrine B Osborne, Nicole Wiedmann

[DESCRIPTION]
This protocol is suitable for preserving tissues for anatomical studies of organs and major pelvic ganglia in adult rats. The protocol is performed under anesthesia and should incorporate all local requirements for standards of animal experimentation.

[STEPS]
SECTION: Preparation for perfusion
1. Make up the following solutions:

Perfusion prewash solution: To 300 ml 0.9% sodium chloride (w/v) add 3.75 ml 1% sodium nitrite (w/v) and 0.11 ml heparin (5000 IU/ml). This is made up immediately prior to use.

Fixative: 4% paraformaldehyde in 0.1M phosphate buffer, pH 7.4. This is made up no longer than 48h prior to use, stored at 4C and brought to room temperature on the day of perfusion.
SECTION: Perfusion
2. Induce anesthesia by an intraperitoneal injection of ketamine (100 mg/kg) and xylazine (10 mg/kg).
SECTION: Perfusion
3. After opening the chest cavity, inject the left ventricle with mixture of 0.25 ml heparin (5000 IU/ml) and 0.5 ml 1% sodium nitrite.
SECTION: Perfusion
4.  A needle connected to tubing (T-connector to prewash and perfusion solutions), and a peristaltic pump (setting: 50 ml/min) is then inserted into the left ventricle and clamped into place using hemostats. Make a small incision in the right atrium to drain blood and perfusate during the procedure.
SECTION: Perfusion
5.  Perfuse with pre-wash solution until the fluid flowing from the right atrium is clear, the liver and extremities are pale (typically 2-3 min). 
SECTION: Perfusion
6. Dissect tissues required for analysis and place in fixative overnight at 4 degrees. 
SECTION: Post-perfusion
7. Wash tissues with phosphate-buffered saline (PBS; 0.1 M, pH7.2), 3 x 30 min washes.
SECTION: Post-perfusion
8. Store in PBS containing 0.1% sodium azide at 4C until used for immunohistochemistry or other microscopic analysis.
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p>The provided data are example raw data acquired with simultaneous multislice imaging (SMS) in a preclinical MRI experiment.</p>
<p>zip includes:</p>
<p> </p>
<p>- Matlab code (Main.m) and needed functions</p>
<p>- data arrays</p>
<p> </p>
<p>Password needed to unzip: smsreconstructionexample</p>
<p> </p>

[GUIDELINES]
<p>- documented example code in dataset -</p>

[STEPS]
?.
TITLE: Ubiquitin immunoprecipitation using an anti-ubiquitin nanobody
AUTHORS: Cole S Sitron, Victoria A Trinkaus, F Ulrich Hartl

[DESCRIPTION]
This protocol describes a method to detect ubiquitination on a protein of interest. This technique relies on immunoprecipitation (IP) of ubiquitinated proteins from a cell lysate through the use of an anti-ubiquitin nanobody coupled to agarose beads. The eluate can be run on SDS-PAGE to determine whether the protein of interest was recovered in the IP and, therefore, ubiquitinated.

[STEPS]
SECTION: Cell collection and lysis
1. Remove the medium from the wells and trypsinize with 500 µL TrypLE Express.

SECTION: Cell collection and lysis
2. Quench the TrypLE Express with 500 µL 10% FBS and move the cells into a centrifuge tube.

SECTION: Cell collection and lysis
3. Pellet cells at 1500 x g for 3 min at 4 °C.

SECTION: Cell collection and lysis
4. Wash cells with 1 mL PBS and pellet again.

SECTION: Cell collection and lysis
5. Resuspend cells in 150 µL Triton Lysis Buffer and incubate on ice for 5 min.

SECTION: Cell collection and lysis
6. Sonicate in the BioRuptor for 5 cycles of 30 s on and 30 s off at 4 °C.

SECTION: Cell collection and lysis
7. Clarify the lysate by centrifugation 18000 x g for 10 min 4 °C.

SECTION: Cell collection and lysis
8. During the centrifugation, prepare BSA standards according to the Rapid Gold BCA Protein Assay Kit manual and begin equilibrating 50 µL of Ubiquitin pan-selector resin (pipette with a cut p200 tip) in 500 µL of lysis buffer.

SECTION: Cell collection and lysis
9. Collect supernatant and place into a new tube on ice.

SECTION: Cell collection and lysis
10. Prepare a small 1:10 dilution of samples and perform Rapid Gold BCA Protein Assay according to manufacturer’s instructions.
SECTION: Immunoprecipitation (IP)
11. Dilute samples into Triton Lysis Buffer in two tubes: 1 mg in 500 µL (for IP) and 100 µg in 25 µL (for input).

SECTION: Immunoprecipitation (IP)
12. Pellet the beads at 1000 x g for 2 min at 4 °C.

SECTION: Immunoprecipitation (IP)
13. Pull off the supernatant, add the 1 mg of lysate to the beads, and incubate with rotation at 4 °C for 60 min.

SECTION: Immunoprecipitation (IP)
14. During the incubation denature the input sample with 25 µL 4X NuPAGE LDS Sample Buffer at 95 °C 5 min.

SECTION: Immunoprecipitation (IP)
15. Pellet the beads at 1000 x g for 2 min at 4 °C.

SECTION: Immunoprecipitation (IP)
16. Wash the beads with either 1 mL Triton Wash Buffer.

SECTION: Immunoprecipitation (IP)
17. Pellet the beads and repeat for a total of 3 washes.
SECTION: Immunoprecipitation (IP)
18. Remove the supernatant and add 100 µL of 2X NuPAGE LDS Sample Buffer.

SECTION: Immunoprecipitation (IP)
19. Elute by boiling at 95 °C for 5 min.

SECTION: Immunoprecipitation (IP)
20. Transfer the beads to a MiniSpin column placed in a tube.
SECTION: Immunoprecipitation (IP)
21. Centrifuge 3000 x g for 3 min.

SECTION: Immunoprecipitation (IP)
22. The eluate has now been collected in the tube and is ready for SDS-PAGE analysis.
TITLE: TRANSFECTION OF i3NEURONS (Support Protocol 3) 
AUTHORS: Michael S. Fernandopulle, Ryan Prestil, Christopher Grunseich, Chao Wang, Li Gan, Michael E. Ward

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><span>Transient protein expression can easily be studied in i</span><span style = "vertical-align:super;">3</span><span>Neurons using lipid-based transfection. This protocol is identical to that in iPSCs (see </span><a href="https://www.protocols.io/view/basic-protocol-2-lipid-mediated-transfection-of-ip-5deg23e" style = "text-decoration:underline;color:blue;cursor:pointer;"><span style = ":;">Basic Protocol 2</span></a><span>). i</span><span style = "vertical-align:super;">3</span><span>Neurons are modestly transfectable, with 5 % to 10 % of cells showing fluorescent protein expression after 24 hr. We have found that refreshing neuronal medium 1 to 2 hr after transfection both allows successful DNA entry into cells and largely prevents cytotoxicity resulting from the transfection reagent. Unlike iPSCs, i</span><span style = "vertical-align:super;">3</span><span>Neurons show increased protein expression/accumulation over time, with greater fluorescence 48 to 72 hr after transfection than at 24 hr. Transient transfections also show more durable expression in i</span><span style = "vertical-align:super;">3</span><span>Neurons than iPSCs, likely because episomes are not diluted by cell division. i</span><span style = "vertical-align:super;">3</span><span>Neurons can be transfected in suspension (i.e., re-plating after day 3 of differentiation) or as an adherent culture, although better results are observed in adherent cultures. They are also amenable to serial transfections (i.e., re-transfecting with the same construct 24 hr apart) if higher-percentage transfections are desired.</span></div></div>

[STEPS]
TITLE: Tissue Sectioning Guidelines - CODEX/PhenoCycler
AUTHORS: Santhosh Sivajothi

[DESCRIPTION]
The purpose of this SOP is to provide general guidelines for preparing tissue sections suitable for PhenoCycler/CODEX

[STEPS]
SECTION: Tissue Sectioning Guidelines for CODEX/PhenoCycler
1. Cut FFPE sections on a microtome at a thickness of 5 microns. Section thickness should not exceed 10 microns.
SECTION: Tissue Sectioning Guidelines for CODEX/PhenoCycler
2. For best results, the tissue should be completely adhered to the slide with minimal tears or folds.
SECTION: Tissue Sectioning Guidelines for CODEX/PhenoCycler
3. Place tissue section or sections within the boundary indicated on the slide below. If tissue overlaps the boundary, proper flowcell seal (coverslip) will not be formed
SECTION: Tissue Sectioning Guidelines for CODEX/PhenoCycler
4. Multiple tissues can be placed within the boundary, as long as there is no overlap of paraffin over adjacent tissues

                                                       

SECTION: Tissue Sectioning Guidelines for CODEX/PhenoCycler
5. Example of good tissue placement: 

            

SECTION: Tissue Sectioning Guidelines for CODEX/PhenoCycler
6. Dry tissue sections overnight at RT
SECTION: Tissue Sectioning Guidelines for CODEX/PhenoCycler
7. Ater drying, store cut slides at 4°C (for up to six months)
SECTION: Tissue Sectioning Guidelines for CODEX/PhenoCycler
8. NOTE: Do not place any stickers or labels on the frosted end of the slide as it may interfere with coverslip adhesion. Use markers or directly print labels on the slide.
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p>The authors describe the skin thickness measurement, based on the analysis of images, obtained using a handheld DermaLab® USB Series ultrasound from Cortex Technology. Epidermis and dermis layers of newborns are obtained by ultrasound scanning. The procedures of the mean thickness determination are pictured.</p>
<p> </p>
<p>We developed a dedicated software to provide epidermal thickness measurement (IP register BR 51 2017 000061 1). The calculation of the epidermal thickness for each line in the image array was based on the size of the pixel in the frame obtaining 356 thickness values. Using a bootstrap technique in the selected sample size with 70 lines, the software resampled the thickness values 2,000 times. From a Gaussian distribution adjustment curve to the epidermis measurement, the peak was the thickness value, and the full with a half-maximum area was the sigma error value. </p>

[BEFORE_START]
<div title="Page 12">
<div>
<div>
<p>Preparation of the probe. </p>
<div title="Page 12">
<div>
<div>
<p>Mounting the water barrier of the probe.</p>
</div>
</div>
</div>
</div>
</div>
</div>

[GUIDELINES]
<div title="Page 3">
<ol>
<li>The skin scanning follows the Instruction Manual of the equipment manufacturer, DERMALAB® SERIES SKINLAB USB, 20 MHz Probe, available in http://www.cortex.dk/dermatology/dermalab-usb-series/.</li>
<li>The newborns are evaluated inside their incubators or in an open heating crib, wherever they were taken care of to ensure minimum manipulation and stable clinical conditions.</li>
<li>Automated measurements of skin layer thickness involved the use of two different software for digital image processing:</li>
</ol>
<ul>
<li>DermaLab® embedded software provided dermal measurements. </li>
<li>For the epidermal axial dimension estimation, we developed a dedicated software in Python language available in the Health Informatics Center of Universidade Federal de Minas Gerais, Brazil: https://site.medicina.ufmg.br/cins/</li>
</ul>
</div>

[STEPS]
?. 
?. 
?. 
?.
TITLE: Transportmedium
AUTHORS: Bettina Ergün

[DESCRIPTION]
Transportmedium für Gewebe aus dem OP bzw. Schnellschnitt.
Das Transportmedium enthält eine erhöhte Konzentration an Zellshield zur Keimreduzierung im Ausgangsmaterial der Zellisolierung.
Das Gewebe wird 1 h bei RT in Transportmedium inkubiert (alternativ: ü. N. 4 °C)

[STEPS]
SECTION: Transportmedium
1.
TITLE: Installation instructions for phylogenetic analysis using a conda environment
AUTHORS: Laise Moraes

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">The purpose of this protocol is to define and provide instructions for creating a conda environment for phylogenetic analysis</div></div>

[STEPS]
?. [Installing Miniconda]
Create a directory called softwares to your HOME directory, switch to it and then download the 64-bit Python 3 Miniconda installer.Install Miniconda quietly, accepting defaults.After installation, remove the Miniconda installer from directory.Set the Miniconda permanent PATH and update conda packages to the latest compatible version.
?. [Creating conda environment]
Create a YML file to build a phylogenetic analysis environment called phy.
?. [Creating conda environment]
Create the environment from the phy.yml file
TITLE: CPEC Protocol
AUTHORS: N.J. Hillson

[STEPS]
?. Measure the DNA concentration (ng/ml) of each assembly piece.
?. Add 100 ng of the linearized vector backbone and equimolar amounts of the other assembly pieces to a 25 ml total volume assembly reaction mixture as follows:linearized vector backbone (100 ng)+ each additional assembly piece (to equimolar with backbone)+ 5 ml 5X HF Phusion Reaction Buffer+ 1 ml 10 mM dNTPs+ 0.75 ml DMSO+ 0.5 ml 2U/ml Phusion Polymerase+ _______dH2O to25 ml
?. Perform the assembly reaction in a thermocycler as follows:30 sec @ 98 C     1 cycle10 sec @ 98 C }30 sec @ 55 C }     1 to 15 cycle(s)**length* (kb) x 15 sec @ 72 C }10 min @ 72 C     1 cycle*The total length of the assembled product (in kb)**The number of repeated cycles should exceed the number of assembly piecesTransform 5 ml of the assembly reaction into 100 ml of competent E. coli and/or run a diagnostic agarose gel to check for successful assembly.
TITLE: Intraperitoneal L-dopa injection in mice
AUTHORS: natalia.lopezgonzalezdelrey, Zachary Gaertner

[DESCRIPTION]
Intraperitoneal L-dopa injection in mice

[STEPS]
SECTION: Preparation
1. Mice were acclimated to the room for 60 min 

SECTION: L-dopa preparation
2. Prepare the L-dopa mix fresh every day.
Prepare a master mix of: 12.5mg/kg L-dopa and 12.5mg/kg benserazide dissolved in sterile saline with 0.25% citrate (wt/vol). Calculate grams depending on number of mice to be injected. Using vortex will dissolve the solution easier.



SECTION: L-dopa injection
3. Mice are injected intraperitoneally with the 12.5mg/kg L-dopa and 12.5mg/kg benserazide solution.
SECTION: Behaviour
4.  One hour post injection, mice were tested using the appropiate behavior protocol.
TITLE: SPARC Serotonin 2A Receptor (5-HT2AR) Immunohistochemistry Protocol in Rat Tissues Labeled with Cholera Toxin B-fragment
AUTHORS: Elisa Gonzalez-Rothi, Yasin Seven, Latoya Allen, Marissa Ciesla, Gordon Mitchell

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">This protocol describes the immunofluorescent labeling technique used to identify serotonin 2A receptor expression in CtB-labelled phrenic motor neurons and within a defined region of interest surrounding phrenic motor neurons.</div></div>

[STEPS]
?. Day 1: primary antibodies required:5-HT2AR: Rabbit anti-5-HT2AR (Immunostar #24288)Cholera toxin B-fragment: Goat anti-CT-B (Millipore #227040)
?. Place 40um transverse spinal cord sections into 1xPBS in 12 well cell culture plates
?. 5x washes in  1xPBS for 5 minutes each at room temperature
?. Blocking: place tissues in 5% Normal Horse Serum (NHS) in 1xPBS-Triton (0.1%) +0.5% bovine serum albumin (BSA) for 60 minutes at room temperature
?. Primary Antibody Incubation: Incubate tissues in: 5%NHS in 1xPBS-Triton (0.1%), Rabbit anti-5-HT2AR (1:1000), and Goat anti-Ct-B (1:2500) for 48 hours at 4 degrees C
?. Day 2: secondary antibodies required:AlexaFluor 488: donkey anti-goat (Invitrogen, Ref#A11055)AlexaFluor 594: donkey anti-rabbit (Invitrogen, Ref#A21207)
?. 5x washes in  1xPBS for 5 minutes each at room temperature
?. Secondary Antibody Incubation: Incubate tissues in: 5%NHS in 1xPBS-Triton (0.1%), donkey anti-goat (1:1000), and donkey anti-rabbit (1:1000) for 2 hours at room temperature
?. 5x washes in  1xPBS for 5 minutes each at room temperature
?. Mount tissues on Superfrost Plus microscope slides
?. Allow slides to dry overnight
?. Coverslip with VectaShield Antifade Hard Set Mounting Medium (Cat#:H-1400)
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
This is a guide to prepare PFA from powder. This is useful for PFA Fixation.

[GUIDELINES]
<span style="text-decoration: underline;">Working Dilutions: </span><br /><span style="font-family: Arial,Helvetica,sans-serif;">Thawed aliquots are stable at 4&ordm; C for up to 2 weeks.<br /></span><span style="font-family: Arial,Helvetica,sans-serif;">Dilute 1 part 2% PFA to 3 parts cells in PBS (eg 60 &micro;l + 180 &micro;l to yield 0.5% final concentration).</span>

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