eamag/protocols-with-steps huggingface dataset:
TITLE: Anthropometry (Standing Height)
AUTHORS: g.spencer

[DESCRIPTION]
Standing height is an assessment of maximum vertical size. Take this measure on all patients aged 6 years and older who are able to stand unassisted. Standing height is measured with a fixed stadiometer with a vertical backboard and a moveable headboard.

[STEPS]
SECTION: Equipment Required
1. 
 

SECTION: Preparing for Measurement
2. Have the patient remove their footwear, hair accessories or hats from the top of their head
SECTION: Standing Height Measurement
3. 1. Ask the patient to stand erect, arms hanging at their sides, feet together, heels, buttocks and back in contact with the stadiometer.

2. Ask the patient to look straight ahead.

3. Ensure the patient’s head is within the Frankfort plane (below)

  
4. Lower the stadiometer headboard and ask the patient to stand as tall as possible and take a deep breath whilst the measurement is being taken.

5. Lower the head plate firmly onto the head with sufficient pressure to compress the hair.

6. Record the height measurement to the nearest 0.5cm.

7. Ask the patient to step down and away from the stadiometer.
SECTION: Considerations
4. Some patients may have conditions that interfere with the specific procedures for measuring stature. One of the more common conditions is kyphosis which is a forward curvature of the spine that appears as a hump or crooked back condition. Kyphosis most frequently occurs in the elderly, and in women the condition is commonly referred to as dowager’s hump. In these cases, it is important to get the best measure possible according to the protocol and to record a “not straight” comment.

If the patient has a hair style which stands well above the top of their head, (or is wearing a turban), bring the head plate down until it touches the hair/turban. With some hairstyles you can compress the hair to touch the head. If you cannot lower the head plate to touch the head and think that this will lead to an unreliable measure, record this. If it is a hairstyle that can be altered, e.g. a bun, if possible, ask the patient to change/undo it.

If the patient is tall, it can be difficult to line up the Frankfort Plane in the way described. When you think that the plane is horizontal, take one step back to check from a short distance that this is the case
TITLE: Protocol for Mouse IP Anesthesia (Ketamine-Xylazine)
AUTHORS: Haowei Li, Markus Holzl, Mohsen Khosravi-Maharlooei, Nichole Danzl, Austin Chen, Megan Sykes

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">This protocol details the constitution of our anesthesia cocktail that we use for sedating our mice when performing surgeries. The surgeries we typically perform are thymectomies and implantation of material under the kidney capsule (through open abdomen or minimally invasive approaches).</div><br/><br/><br/><br/></div>

[STEPS]
?. [For Intraperitoneal (IP) Approach:]
Use 10 uL/gram of anesthesiaComments:1. It takes time until the mice fall asleep 2. Very often, mice do not fall asleep but move around like zombies. This requires another small dose of IP ketamine-xylazine injection or a small dose of IV injectiona. When doing IP injection again, we have injected IP with a volume of about 1/3 of original volumeb.  For IV approach after IP, it takes very little anesthesia for them to fall asleep. We usually use around 10-20 uL. Inject this slowly into the mouse. The mouse should fall asleep rapidly. It is very easy to overdose with IV injection so be careful.
TITLE: GENERATION OF LYSATE INOCULATION MATERIALS
AUTHORS: Scott Vermilyea

[DESCRIPTION]
This protocol details inoculation lysate preparation.

[STEPS]
SECTION: Soluble/Insoluble lysate preparation from harvested brainstem/spinal cord tissue
1. Acquire the tissue  from 4-month-old asymptomatic TgA53T (Line G2-3) and end-stage tissue harvest and store at -80 °C prior to use.

SECTION: Soluble/Insoluble lysate preparation from harvested brainstem/spinal cord tissue
2. Weigh the tissue and suspend it in 0.9% sterile saline (1:10 w/vol).
SECTION: Soluble/Insoluble lysate preparation from harvested brainstem/spinal cord tissue
3. Homogenize and centrifuge for 5 min at 3000 x g at 4 °C. The supernatant (S3000) is obtained.



SECTION: Soluble/Insoluble lysate preparation from harvested brainstem/spinal cord tissue
4. Centrifuge S3000 for 45 min at 150000 x g at 4 °C. The supernatant (S150) – Highly soluble fraction is obtained.

SECTION: Soluble/Insoluble lysate preparation from harvested brainstem/spinal cord tissue
5. Wash pellet and resuspend in sterile saline (half of original volume) by sonication (3 x 10 min pulses). Resuspended pellet (P150) – Insoluble fraction is obtained.


SECTION: Endoplasmic Reticulum (ER)-enriched fractionation
6. Homogenize freshly harvested tissues (1:10 w/vol) in lysis buffer (refer material section).
SECTION: Endoplasmic Reticulum (ER)-enriched fractionation
7. Centrifuge at 1000 x g and collect the supernatant.

SECTION: Endoplasmic Reticulum (ER)-enriched fractionation
8. Centrifuge at 10000 x g. The pellet containing mitochondria is obtained.

SECTION: Endoplasmic Reticulum (ER)-enriched fractionation
9. Centrifuge supernatant at 100000 x g. The supernatant containing cytosol and pellet containing ER are obtained.

SECTION: Soluble/Insoluble lysate preparation from harvested brainstem/spinal cord tissue
5.1. Wash pellet and resuspend in sterile saline (half of original volume) by sonication for 10 s pulses) (1/3).
SECTION: Soluble/Insoluble lysate preparation from harvested brainstem/spinal cord tissue
5.2. Wash pellet and resuspend in sterile saline (half of original volume) by sonication for 10 s pulses) (2/3).
SECTION: Soluble/Insoluble lysate preparation from harvested brainstem/spinal cord tissue
5.3. Wash pellet and resuspend in sterile saline (half of original volume) by sonication for 10 s pulses) (3/3).
TITLE: Obtaining Competent Cells
AUTHORS: Miquel Estévez-Gay

[DESCRIPTION]
Obtencion of 100ml competent E.coli aliquots that must be stored in -80C 

[GUIDELINES]
Competent cells are fragile. All work must be done under a laminar flux cabin and in aseptic conditions.

[STEPS]
1. Incubate 10ml of E.coli BL21(DE3) or DH5α o/n without plasmid (from –80°C stock). To do that, add 10 mL of sterile LB media in a 50ml sterile falcon tube. With the use of a yellow micropipette, scratch the surface of the frozen E.coli from the -80°C freezer and toss it into the Falcon containing the media. This must be done fast, and the E.coli must not be outside the freezer more than 1 or 2 minutes (Use the termoblock from the -20°C to keep the E.coli stock frozen).

2. Incubate      

3. Re-inoculate in a new 10 mL LB. Incubate until absorbance is 0.6-0.7 OD550 120 min.

4. Centrifuge 4000 rpm, 10 min, 4 °C

5. Under the Laminar flux Cabin, throw the supernatant and resuspend in the same volume of CaCl2 100 millimolar (mM)cold and sterile. Do not use vortex! 


6. Incubate in ice 30 min. 

7. Centrifuge again 4000 rpm, 10 min, 4 °C

8. Resuspend in 1/10th of the previous volume of cold and sterile CaCl2 and 15% glycerol. Aliquot them in 100 µL (using 1.5ml centrifuge tubes).

9. These aliquots can be used right away or can be stored in the -80°C freezer.
TITLE: Thawing Rosettes and NPC
AUTHORS: Celeste Karch, Rita Martinez, Jacob Marsh

[STEPS]
?. Aliquot  of pre-warmed DMEM/F12 into a 15 ml conical tube.
9 ml
?. Remove cells from liquid nitrogen and thaw in water bath for approximately  or until a small ice clump remains.
?. Add freshly thawed cells into theof pre-warmed DMEM/F12 and mix gently by gently shaking tube 3 times. Avoid breaking up clumps of cells.
9 ml
?. Centrifuge cells at 750 rpm for . Aspirate supernatant.
?. Resuspend in  of NIM and plate into one well of a 6-well plate.
2 ml
If cell viability count is necessary, after resuspension remove 10 μl into a separate microfuge tube for cell viability count .
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p>Using UV-induced peptide exchange, MHC/peptide monomers can be generated with conditional Flex-T™ monomers that harbor peptides of interest in their binding grooves. These new MHC monomers are subsequently multimerized using streptavidin-fluorophore conjugates. The resulting Flex-T™ reagents can be used for staining antigen-specific T cells and flow cytometric analysis. In humans, the MHC molecules are called HLA (Human Leukocyte Antigen).</p>

[GUIDELINES]
<p><strong>Materials</strong><br />- Phosphate buffered saline pH 7.4, 10X concentrate (PBS, BioLegend Cat# 926201)<br />- Conditional Flex-T™ monomers<br />- 10 mM peptide solution of choice in 100% DMSO<br />- DMSO (e.g. Sigma-Aldrich Cat#D5879)<br />- 50 mM D-Biotin (e.g. Thermo Fisher, Cat#B20656)<br />- 10% (w/v) NaN3 (e.g Sigma, Cat#S2002)<br />- Fluorophore-conjugated Streptavidin (BioLegend Cat# 405203, Cat# 405207, Cat#405225 or equivalent)<br />- Cell Staining Buffer (BioLegend Cat#420201 or equivalent)<br />- 96-well Polystyrene Microplate, U-shape (e.g. Falcon Cat#353077) or 5mL, 12 x 75mm tubes (e.g. Falcon Cat# 352008)<br />- Plate sealers (BioLegend Cat# 423601)<br />- 1.5 mL tubes (e.g. Eppendorf Cat# 022364111)</p>
<p> </p>
<p><strong>Equipment</strong><br />- UV lamp, long-wave UV, 366 nm, 8 Watts (For example CAMAG cat# 022.9115, or Ultraviolet Crosslinker CL-1000)<br />- Incubator (37°C)<br />- Centrifuge capable of accommodating microtiter plates and tubes<br />- Single and multichannel pipettes capable of accurate delivery of variable volumes, and pipette tips</p>
<p> </p>
<p><strong>Precautions for use</strong><br />- DMSO can be used to dissolve the peptides. However, do not exceed an end concentration of 10% (v/v) in the exchange reaction.<br />- Avoid repeated freeze-thawing.<br />- The Flex-T™/peptide solution needs to be kept on ice in the dark as much as possible. Do not work in front of a window.<br />- The use of short-wavelength (254 nm) or broad-band UV lamps is detrimental to MHC complexes.<br />- Centrifuge all vials before use (1 minute 3000xg at 4°C).</p>
<p> </p>
<p><strong>View protocol on Biolegend website for Representative Data</strong></p>

[STEPS]
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TITLE: Green Lab Nanoparticle For 6 Well Cardiomyocyte Transfection    
AUTHORS: Edwin Yoo

[DESCRIPTION]
nanoparticle transfection    

[STEPS]
SECTION: Nanoparticle Synthesis
1. DNA Dilution
For 3 replicate wells of one condition in 6-well plate format, mix the following in microcentrifuge tube.
60 ul DNA at 1ug/ul
440 ul sodium acetate
Total volume = 500 ul for 3 replicate wells per one condition
SECTION: Nanoparticle Synthesis
2. Polymer Dilution (7.2 mg.ml polymer concentration in NaAc)
For 3 replicate wells of one condition in 6-well plate format, mix the following in microcentrifuge tube.
40 ul polymer (e.g. 4-5-6)    
460 ul sodium acetate
Total volume = 500 ul for 3 replicate wells per one condition 
SECTION: Nanoparticle Synthesis
3. Nanoparticle Synthesis
Combine diluted DNA with diluted polymer
Add 500 ul of diluted DNA to 500 ul diluted polymer (can freeze here at -80 at your own risk)
Add 5000 ul of media to 1000 ul NP's for final volume master mix of 6000 ul per condition
Incubate for 10 minutes
SECTION: Nanoparticle Synthesis
4. Transfection
Change media from each well of 6 well plate with 2000 ul nanoparticle master mix (media + nanoparticles)
Incubate for 24 hours or desired time
TITLE: 0.5 M EDTA (0.5 L)
AUTHORS: Mattia Pegoraro

[DESCRIPTION]
<p>Is used in a multitude of experiments, but is often used as an ingredient in 10x TBE buffer</p>

[STEPS]
?. Fill 0.5 L beker with of deionized water.
300 ml
?. Add of Ethylenediamine Tetraacetic Acid Disodium Salt (292,24 g/mol) to the water and mix.
73 g
?. Add NaOH pellets until the pH is near 7.8 (the solution will be clear around pH 7.5). Complete to arrive at pH 8 with NaOH 5 M solution (also NaOH 10 M is good).
?. Once at pH 8, fill to.
500 ml
?. Autoclave at for or filter it with 0,22 u filter..
?. Put the electrode pH meter into the solution (with continuos mixing) and mesure the pH (don't worry if the solution isn't clear). The pH will be about 3 at this point. The pH needs to be at 8.
TITLE: Electrophysiology with iPSC-derived neurons
AUTHORS: Minee-Liane Choi

[DESCRIPTION]
This protocol describes the method to perform patch-clamp recordings of iPSC-derived neurons. 

[STEPS]
1. Patch-clamp recordings of iPSC-derived neurons are performed using an infrared differential interference contrast (DIC) imaging system on an Olympus BX51WI upright microscope (Olympus, Japan) coupled with a Multipatch 700B amplifier under the control of pClamp 10.2 software package (Molecular Devices, USA)
2. For the recordings, a neuronal culture or co-culture is plated on glass coverslips, placed in a recording chamber mounted on the microscope stage and constantly perfused with a physiological buffer medium. 
3. Whole-cell recordings are performed using glass pipettes with a resistance of 3.5-6 MΩ
when filled with the intracellular solution. 

 

4. Whole-cell recordings are performed using glass pipettes with a resistance of 3.5-6 MΩ when filled with the intracellular solution. 
 

5. In the whole-cell (immediately after membrane breakthrough), iPSC-derived neurons are recorded
for the resting membrane potential (Vrest), membrane capacitance (Cm), the membrane time constant (τm), and input resistance (Rin)
6. To induce neuronal firing, a series of sub- and supra-threshold rectangular current pulses are applied with a stepwise-increased stimulus intensity at the Vhold set at −60 mV to −70 mV. 

 

7. The analysis of the AP waveform was performed for the first AP only to quantify the
threshold value, the spike amplitude, overshoot, the spike width (duration at
half-maximal amplitude), the rates of depolarisation and re-polarisation phase.
TITLE: Slice Culture Media
AUTHORS: Allen Institute for Brain Science

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">This protocol describes the preparation of Slice Prep Media used for human or mouse slice culture.</div><div class = "text-block"><span style = "font-weight:bold;">Note</span><span>: Research reported in this publication was supported by the National Institute Of Mental Health of the National Institutes of Health under Award Number U01MH114812. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.</span></div></div>

[STEPS]
TITLE: Cryo-EM sample preparation for full length LRRK2(I2020T):MLi-2/GZD-824-E11 DARPin complex
AUTHORS: Marta Sanz Murillo, Amalia Villagran Suarez

[DESCRIPTION]
Guide for cryo-EM grid preparation of FL-LRRK2 bound to Type-I/Type-II inhibitors

[BEFORE_START]
Before starting cryo-EM grid preparation section: Set up Vitrobot at 95% humidity and 4C. Put new paper blot

[STEPS]
SECTION: Protein purification
1. His6-Z-TEV-LRRK2 was expressed and purified as described in a previous protocol

 

SECTION: Sample preparation
2. Prepare LRRK2 buffer. Keep it at 4ºC.
20 millimolar (mM) HEPES pH=7.4
150 millimolar (mM) NaCl
2.5 millimolar (mM) MgCl2
20 micromolar (µM) GDP
0.5 millimolar (mM) TCEP
SECTION: Sample preparation
3. Spin down purified LRRK2 (10000 rcf, 4°C, 10 minutes). Leave protein on ice afterward.
For the best result, keep protein on ice and reduce the amount of time between spinning
and freezing cryo-EM samples.
SECTION: Sample preparation
4. Thaw E11 DARPin and spin it down. Measure its concentration.
SECTION: Sample preparation
5. Dilute inhibitors (diluted in 100% DMSO) to an intermediate desired concentration using LRRK2 buffer.
SECTION: Sample preparation
6. Based on the initial and final LRRK2 concentration, calculate the necessary volume of E11 DARPin to get a proportional ratio LRRK2:E11 DARPin 1:1.25 in the sample. After that, add either MLi-2 or GZD-824. Dilute to your final desired LRRK2 concentration using LRRK2 buffer (150 mM NaCl).
 

SECTION: Sample preparation
7. Incubate 10 minutes at RT. Afterward, keep it on ice until grid preparation.
SECTION: cryo-EM grid preparation
8. We used UltraAuFoil Holey Gold 2/2 200 mesh grids and plasma cleaning them in the Solarus II (Gatan) using the QuantiFoil Au preset.
SECTION: cryo-EM grid preparation
9. Apply 3 to 3.5 microliters (µl) of sample and plunge freeze. We used a
Vitrobot (FEI) to blot away excess sample and plunge freeze in ethane liquid. (In
our case, we use 4 seconds as a time blot as 20 sec as a wait time and 4 as a blot force, but these
parameters are slightly different from one Vitrobot to another. I would try
with the Vitrobot parameters already tested in your machine first).
SECTION: cryo-EM grid preparation
10. Store grids in liquid nitrogen until ready for imaging
TITLE: Apomorphine-induced rotations
AUTHORS: Santiago Unda, Mihaela Stavarache, Michael G. Kaplitt, Roberta Marongiu

[DESCRIPTION]
To test for 6-OHDA lesion efficiency, rotational behaviors are assessed while mice are under the effects of apomorphine. This test is run during the dark phase of light cycle. Experiments are performed by an examiner blinded to treatment groups. 

[STEPS]
2. Prepare apomorphine in 0.9% saline/0.1% ascorbate solution to dose each mouse with 0.25mg/kg via subcutaneous injection.
*Note that this solution is light-sensitive and should be shielded from light.
SECTION: Preparation
1. Weigh mice and habituate to the testing room for at least 1 hour prior to the start of testing.
3. Clean open field arenas.
SECTION: Testing
5. Test mice in groups based on the number of open field arenas available. Before testing each group, start the tracking software. The trial time should not start until the animals are placed inside the arena.
6. For each group, inject each mouse with 0.25 mg/kg apomorphine via subcutaneous injection, and immediately place in the center of the corresponding open field arena. Repeat for the other mice in the same group.
4. Set up tracking software on computer to record trials of 45 min starting once an animal is detected within the arena. Using Ethovision, the automatic animal detection with deep learning settings is used to track center to-nose and center to-tail points in the defined arenas.

SECTION: Analysis
10. Use the video recordings to analyze and count the numbers of rotations >50 degrees clockwise as well as and counterclockwise. 
7. Run the trial for the 45 min period.

8. Remove mice and place back into cages.
9. Clean open field arenas, reset the tracking software for the next trial, and repeat steps for the rest of the groups.
TITLE: Tranfection of sgRNA using SpCas9 containing plasmids to generate cell lines with Cas9
AUTHORS: Binnypreet Kaur, Drahomíra Faktorová,  Julius Lukeš,  ,  

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><div class = "justify" style = "text-align:left">This protocol describes transfection of  ribonucleoprotein (RNP) complexes that consist of purified Cas9 nuclease duplexed with synthetic guide RNA in cultured cells using the AMAXA Nucleo AMAXA Nucleofector II Electroporation system using  Program X-001.</div></div><div class = "text-block"><div class = "justify" style = "text-align:left">RNPs complex have only transient expression inside the cell, this allows for the highest levels of editing efficiency and greatly reduces the chances of possible off target and toxic effects of integration of foreignDNA in host genome</div></div></div>

[STEPS]
?. Step1: Assemble RNP complexes with (30pmol/μl) of gRNA and (20pmol/μl) of Cas9 in the  ratio of (Cas9:gRNA) 1:9  in resuspension buffer  before electroporation and keep it at room temperature for 10 min and then transfer it to ice untill use.
?. Step2: Follow the protocol 7 for transfection
TITLE: Molecular Cloning
AUTHORS: Lizhen Zhu, Ruixin Lin, Xiaoqi Ye, Ruiyan zeng, 18814306949 Minkun Huang, 624787660

[DESCRIPTION]
SZPT-CHINA Experimental methods of molecular cloning

[GUIDELINES]
advisers WeiJia Wu
LiJun Zhang（PhD)
Yongjun Tang(PhD)

[STEPS]
SECTION: Vector linearization and target fragment preparation
1. Q5 DNA polymerase PCR system(50 µL )
 25 µL 
2 µL 
2 µL 
1 µL 
20 µL 
 
PCR Program
  



SECTION: Enzyme digestion Connection
2.  Enzyme digestion Connection 
SECTION: Transformation
3. Transformation
SECTION: Transformation
3.1. Take 50 µL of competent cells in a sterile Eppendorf tube.
SECTION: Transformation
3.2. Add 2~5 µL of the plasmid to be transformed into each tube, gently mix the plasmid and competent cells with a pipette tip, and ice bath for 25 minutes.
SECTION: Transformation
3.3. Heat shock the ice-bath mixture in a 42℃ water bath for 45s, do not shake the centrifuge tube.
SECTION: Transformation
3.4. Take out the ice bath and cool for 2 minutes.
SECTION: Transformation
3.5. Add 800 µL of non-resistant LB medium preheated to 37°C to each tube, shake gently at 37°C, 150r/min for 1h.
SECTION: Transformation
3.6. Centrifugally concentrate to 100μL, spread it on an LB plate containing antibiotics, and incubate inverted at 37°C for 12-16h.
SECTION: Transformation
3.7. Make a negative control at the same time (use distilled water instead of plasmid).
SECTION: Colony PCR screening of positive clones
4. Colony PCR screening of positive clones
SECTION: Colony PCR screening of positive clones
4.1. Add 25ul of sterile water to the 96-well plate, pick about 15-20 colonies from the successfully transformed DH5α bacterial plate with a pipette tip, and mix them evenly. (Make a corresponding mark on the picking plate).
SECTION: Colony PCR screening of positive clones
4.2. Put the mixed bacteria solution into 10ul to 200ul PCR tube and mark it.
SECTION: Colony PCR screening of positive clones
4.3. Take 0.1ul of bacterial solution in a 96-well plate and add it to the antibiotic-containing plate with a pipette tip and mark it.
SECTION: Colony PCR screening of positive clones
4.4. After mixing, the bacteria solution is used for PCR, the system is as follows(20 µL )
5 µL 
0.5 µL 
0.5 µL 
2 µL
2 µL 

The procedure is as follows

  

SECTION: Colony PCR screening of positive clones
4.5. Verification result of 1% agarose gel electrophoresis.
SECTION: Enzyme digestion Connection
2.1. Enzyme digestion and connection system(10 µL )
3 µL 
1 µL 
1 µL 
0.5 µL 
4.5 µL 

SECTION: Enzyme digestion Connection
2.2. Connection system
  

5.
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
Zonulin is a protein, synthesized in intestinal and liver cells, that reversibly regulates intestinal permeability. Zonulin modulates the permeability of tight junctions between cells of the wall of the digestive tract. It was discovered in 2000 by Alessio Fasano and his team at the university of maryland school of medicine. The classic symptom of cholera is profuse, watery, debilitating diarrhea.

[STEPS]
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p>ELISPOT allows detection of single IFN-γ-secreting cell, but the use of unseparated spleen cells removes the advantages of the method due to a high back ground of IFN-γ-production and uncertainty of the phenotype of secreting cells (CD4, CD8 or NK).</p>
<p>We used high purity sorted CD4 and CD8 cells.</p>
<p>For in vitro restimulation of CD8 T-cells, we used dendritic cells that present antigenic epitopes  of  M2,  NP  or  of  adenovirus  in  the  context  of  MHC  class  I.  In  this  instance,  dendritic  cells  were  transduced  with  Ad5-tet-M2,  Ad5-tet-NP  or  Ad5-null, respectively.</p>
<p>For restimulation of CD4 T-cells in vitro, we used dendritic cells that present antigenic M2 or NP epitopes in the context of MHC class II. For this purpose, dendritic cells were additionally activated by lipopolysaccharide E. coli (1 μg/ml) and were loaded with recombinant protein NP or M2e peptide..</p>

[GUIDELINES]
<p> </p>
<p> </p>
<p>Ref.</p>
<p>1. <a href="http://onlinelibrary.wiley.com/doi/10.1002/0471142735.im0301s39/abstract" target="_blank">http://onlinelibrary.wiley.com./doi/10.1002/0471142735.im0301s39/pdf#</a></p>
<p>2. example of sorting https://community.cytobank.org/cytobank/experiments/66457</p>
<p>3. DOI: http://dx.doi.org/10.14440/jbm.2014.12</p>
<p>4. http://www.bdbiosciences.com/ds/ab/others/552569_552138_Book_Website.pdf</p>

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