eamag/protocols-with-steps huggingface dataset:
TITLE: Discharge of LiPo Batteries Using Saltwater Electrolysis 
AUTHORS: marshall.bennett

[DESCRIPTION]
Procedure developed using information gathered from the following sources: 
How To Dispose Of Lipo Batteries? - Standard Battery (standardbatteryinc.com)

Discharging & Disposing of LiPo Batteries – HeliDirect Support Center (zendesk.com)

https://www.researchgate.net/profile/Alireza-Bazargan/publication/352042531_Discharge_of_lithium-ion_batteries_in_salt_solutions_for_safer_storage_transport_and_resource_recovery/links/61ebe8f78d338833e3895d56/Discharge-of-lithium-ion-batteries-in-salt-solutions-for-safer-storage-transport-and-resource-recovery.pdf

[STEPS]
1. Place the battery into a container that is sealable with a vented lid, such as a five gallon bucket. 
3. Carefully pour the salt/water solution into the container holding the battery so that it is completely submerged and covered in several inches of water. 
4. Check for activity from the battery and cover the bucket with the vented lid. Bubbles should emerge from the battery's terminal port as electrolysis begins. 
5. Monitor the battery and the lid regularly. It is possible that residue may develop along the surface of the salt/water solution, and it may take several changes of the water for the battery to completely discharge. Complete discharge of the battery could take many days, depending on the size and build of the battery. 
6. Battery is safe to be recycled or disposed of when electrolysis is complete and voltage is at a low level. Store the battery in a sealable plastic bag until it is ready to be disposed of. 
2. Dissolve Non-Iodized Salt into Water in an appropriate container following the measurements outlined in the chart below.
TITLE: No Title
AUTHORS: 

[GUIDELINES]
<p><strong>Virtualization must be enabled for Docker to work.</strong> This is performed through the BIOS. To access the BIOS you must restart the computer and press the appropriate key immediately upon start up. Each computer has it's own key to enter the BIOS so google you specific machines key. There should be 2-4 different menus. Scroll until you find an option called 'virtualization'. Enable virtualization. Save and Quit BIOS. Docker should be able to run now. </p>
<p> </p>
<p><strong>Data binding for Windows Users:</strong></p>
<p> </p>
<p>The following is a list of files you need to SCP. Remember you need to open a new terminal window in Cygwin and perform these commands offline (meaning don't login to the hpc on the new window):</p>
<p> </p>
<ul>
<li>scp netid@sftp.hpc.arizona.edu:/rsgrps/bh_class/username/assembly/megahit-out/fixed-contigs.fa contigs.fa</li>
<li>scp netid@sftp.hpc.arizona.edu:/rsgrps/bh_class/username/read_recruit/bam/*.bam .</li>
</ul>
<p>The default location for these files is C:\cygwin64\home\USER. You can navigate there and ensure all necessary files are present using File Explorer. This PC &gt; C &gt; cygwin64/home/user</p>
<p> </p>
<p>Move the necessary files to a folder in Documents. </p>
<p> </p>
<p><strong>Starting your Anvi'o session with necessary data:</strong></p>
<p> </p>
<p> </p>
<p>In order to access your data in the Docker image you must execute this command:</p>
<p> </p>
<pre><code class="language-bash" data-lang="bash">docker run --rm -v /path/to/files:/my_data -it meren/anvio:latest<br /><br /></code>where /path/to/files points to where you contig and bam files are located. An easy way to get this is to navigate to the folder containing your files inside the Docker Terminal. pwd to get the path.<br /><strong>Note:</strong> Use / instead of the windows convention of using \ <br />:/my_data is the name of the directory that will be created inside the Docker Image. You can leave my_data as is and just edit the path.<br /><br /><br /></pre>
<p> </p>

[STEPS]
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?.
TITLE: Protocols for "The genome analysis of Tripterygium wilfordii reveals TwCYP712K1 and TwCYP712K2 responsible for oxidation of friedelin in celastrol biosynthesis pathway"
AUTHORS: Tianlin Pei, Mengxiao Yan, Yu Kong, Hang Fan, Jie Liu, Mengying Cui, Yumin Fang, Binjie Ge, Jun Yang, Qing Zhao

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><span style = "font-style:italic;">Tripterygium wilfordii</span><span> is a vine from the Celastraceae family that is used in traditional Chinese medicine (TCM). The active ingredient celastrol is a friedelane-type pentacyclic triterpenoid with putative roles as antitumor, immunosuppressive, and antiobesity agent. Here, we report a reference genome assembly of </span><span style = "font-style:italic;">T. wilfordii</span><span> with high-quality annotation by using a hybrid sequencing strategy, obtaining a 340.12 Mb total genome size, a contig N50 reaching 3.09 Mb. We successfully anchored 91.02% of the sequences into 23 pseudochromosomes using Hi-C technology and the superscaffold N50 reached 13.03 Mb. In addition, we annotated 31,593 structural genes, and a repeat percentage of 44.31%. Comparative evolutionary analyses showed that </span><span style = "font-style:italic;">T. wilfordii </span><span>diverged from species of Malpighiales approximately 102.4 million years ago. Based on integration of the genome, transcriptome and metabolite analyses, as well as </span><span style = "font-style:italic;">in vivo</span><span> and </span><span style = "font-style:italic;">in vitro</span><span> enzyme assays of the two CYP450 genes</span><span style = "font-style:italic;"> TwCYP712K1</span><span> and </span><span style = "font-style:italic;">TwCYP712K2</span><span> the second biosynthesis step of celastrol was investigated. Syntenic analysis revealed that </span><span style = "font-style:italic;">TwCYP712K</span><span>1 and </span><span style = "font-style:italic;">TwCYP712K2</span><span> were derived from a common ancestor. These results provide insights for further investigating pathways related to celastrol andvaluable information to aid the conservation of resources. In addition, they helped us understand the evolution of Celastrales.</span></div></div>

[STEPS]
TITLE: iPSC Transduction
AUTHORS: Ali Albalakhi, Ning Xia

[DESCRIPTION]
This is the iPSC transduction protocol.

[GUIDELINES]
Note: 
Cells will need to be maintained in StemFlex medium to reform healthy iPSCs colony-like morphology (4-5 days) 
Given that we don’t know what viral titer will be toxic to the cells during transduction we will have to optimize the transduction as we progress 

[STEPS]
SECTION: Begin transduction with cell line 7026
1. 
SECTION: Day 1:
2. Split iPS cells into single cells in complete medium with RVC. (500µl per well)
SECTION: Day 1:
3. Seed cells at 5.0X104 cells in glass-bottom 24-well plates
SECTION: Day 2: Calculations are for 24-well plates. (If using a 6 well plate, then seed at 1x105 cells per well, 850µL of medium the next day with 150µL)
4. Virus infection:
SECTION: Day 2: Calculations are for 24-well plates. (If using a 6 well plate, then seed at 1x105 cells per well, 850µL of medium the next day with 150µL)
5. Remove the culture medium from cells, add (300µL) fresh medium without RVC into each well

SECTION: Day 2: Calculations are for 24-well plates. (If using a 6 well plate, then seed at 1x105 cells per well, 850µL of medium the next day with 150µL)
6. Add 45µl virus and 1µg/ml polybrene into cells and mix it. Incubate in 37 degree, 5% CO2.



 


SECTION: Day 2: Calculations are for 24-well plates. (If using a 6 well plate, then seed at 1x105 cells per well, 850µL of medium the next day with 150µL)
7. Add Polybrene into the infected well to make the final concentration to be 1ug/ml.
SECTION: Day 2: Calculations are for 24-well plates. (If using a 6 well plate, then seed at 1x105 cells per well, 850µL of medium the next day with 150µL)
7.1. Stock Polybrene (10mg/mL) 
SECTION: Day 2: Calculations are for 24-well plates. (If using a 6 well plate, then seed at 1x105 cells per well, 850µL of medium the next day with 150µL)
7.2. Make a 100x dilution to make a working concentration of 100µg/mL

(10mg/mL) (Vstock) = (100µg/mL)(0.5mL) 
Vstock = 5µL of stock into 495µL of DPBS (-)(-)

(100µg/mL) (Vstock) = (1µg/mL)(0.3mL) --> Vstock = 3µLof working concentration per 300µL of medium added 
SECTION: Day 3:
8. Monitor the cells for the next 48 hours 
SECTION: Day 4 (or 48 hours later):
9. Check IRES-GFP or HLA-A2’s expression at least 48 hours after the transduction. 
SECTION: Day 4 (or 48 hours later):
10. For antibiotic-based selection, the cells will be maintained to reform colonies and subsequently grown in medium with 1µg/mLof Puromycin to initiate selection
TITLE: Purification of total RNA (microRNA and mRNA) from liquid samples
AUTHORS: atsuko mizoguchi

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Total RNA was extracted from cells and the cultured supernatant and exosome which was extracted from the same amount of supernatant using 3D-Gene® RNA extraction reagent.Comprehensive miRNA and mRNA expression analysis were performed using 3D-Gene® Human miRNA Oligo Chip (miRBase ver.21) and mRNA Oligo Chip(Toray Industries, Inc.) which are featured with the columnar structure and bead-mixing for high sensitivity.</div></div>

[STEPS]
?. For 300ul of cultured supernatant and exosome in 1.7ml centrifuge tube, add 900 μl 3D-Gene RNA extraction  (Toray Industries,Inc., Kanagawa, Japan)reagent from a liquid sample kit.
?. Homogenize by vortexing for 1min to mix.Place the tube containing the homogenate on the benchtop
[incubation]
?. [incubation]
?. Centrifuge for 10 min at 12,000 x g at 4°C without brakes. After centrifugation, heat the centrifuge up to room temperature.
[incubation]
?. Transfer the 300ul of upper aqueous phase to a new collection tube.
?. Add 450ul of 100% ethanol and mix thoroughly by pipetting up and down several times. Do not centrifuge.
?. Transfer the 750ul of sample into the silica-based membrane column and purification according to general nucleic acid purification protocol.
?. Elute the RNA with RNase free water, and concentrate to 4ul.
?. Comprehensive miRNA and mRNA expression analysis were performed using 3D-Gene® Human miRNA Oligo Chip (miRBase ver.21) and mRNA Oligo Chip(Toray Industries, Inc.) which are featured with the columnar structure and bead-mixing for high sensitivity.
TITLE: Bionano genome mapping from animal tissue
AUTHORS: Graham J Etherington, Darren Heavens, David Baker, Ashleigh Lister, Rose McNelly, Gonzalo Garcia, Bernardo Clavijo, Iain Macaulay, Wilfried Haerty, Federica Di Palma

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Bionano genome map protocol</div></div>

[STEPS]
?. The Bionano genome maps were prepared by taking approximately  of DNA (in this case extracted from a polecat) from the sample stored in 100% EtOH.
10 mg
?. The IrysPrep Animal Tissue DNA Isolation from Fibrous Tissue protocol was followed using the IrysPrep Animal Tissue DNA kit (RE-013-10) from Bionano Genomics.
?. The animal tissue sample was cut up into  before blending using a Qiagen Tissueruptor.
on ice
?. Spin at 1500x g for  in centrifuge, remove supernatant and resuspend in around  of HB buffer using wide bore tip to produce a total volume of .
50 µl
66 µl
?. of LMP agarose was then melted at  and cooled to  before addition to the cell resuspension and mixing using a wide bore tip.
40 µl
70 °C
43 °C
?. One plug of around  was cast using the Chef Mammalian Genomic DNA Plug Kit (Bio-Rad 170-3591).
90 µl
?. Once cooled to  the plug was added to a lysis solution containing  proteinase K (QIAGEN 158920) and  of Bionano lysis Buffer.
4 °C
200 µl
2.5 ml
?. This was incubated at  for  in a thermomixer, making a fresh proteinase K solution and incubating .
50 °C
?. The  tubes were then removed from the thermomixer for  before  RNAse A (Qiagen158924) was added and to the tubes, returned to the thermomixer for a further hour at .
50 ml
50 µl
37 °C
?. The plugs were then washed 7 times in the Wash Buffer supplied with the Chef kit and 7 times in 1xTE.
?. The plug was removed and melted for  at  followed by  at  before adding  of 0.2U/µl of GELase (Cambio Ltd G31200).
70 °C
43 °C
10 µl
?. After  at  the melted plug was dialysed on a  membrane (Millipore VCWP04700) sitting in  of 1xTE buffer in a small petri dish.
43 °C
0.1
15 ml
?. After  the sample was removed with a wide bore tip and mixed gently  and left  at . A small amount was removed to QC on an Opgen Argus Q-Card and Qubit HS to calculate the DNA concentration.
4 °C
?. of DNA was taken into the NLRS (Nick, Label, Repair and Stain) reaction using  Nt.BspQI (NEB R0644S). The optical maps were then generated using the Bionano Irys platform.
300 ng
1 µl
TITLE: Preparation of JF dye for retro-orbital injection in mice
AUTHORS: Boaz Mohar

[DESCRIPTION]
How to prepare JF dyes for retro orbital injection.
 

[STEPS]
SECTION: Dye preperation
1. Take an aliqote with 100 nmol  of dye from -20 °C storage.


SECTION: Dye preperation
2. Add 20 µL  DMSO and do ~10 up and down with the pipeter


 


SECTION: Dye preperation
3. Add 20ul Pluronic F-127 20% in DMSO ~10 up and down with the pipeter


 


SECTION: Dye preperation
4. Add 60ul 1x PBS ~10 up and down with the pipeter
 

SECTION: Dye preperation
5. Drew up to 100ul without bubbles to a 29G syringe
 



SECTION: Dye preperation
6. Follow the Retro-orbital injection protocol
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p><strong><em>Gene expression</em></strong> - <strong><em>MQRT-PCR</em></strong></p>
<p>Reverse transcriptase reactions were performed using High Capacity cDNA Reverse transcription kit (Applied Biosystems, FosterCity, CA, USA) according to the manufacturer ’ s protocol with 350 ng of total RNA. Multiplex quantitative RT-PCR Real-time PCR was performed using Rotor-Gene Q (Qiagen, Hilden, Germany). All reactions were run in triplicate using PerfeCTa® MultiPlex qPCR SuperMix (Quanta Biosciences, Inc. Gaithersburg, MD, USA) with primer and probe sets for target genes at 300 nM concentration each. 3 ′ 6-Carboxyfluorescein (FAM), 6-Hexachlorofluorescein (HEX), Texas Red and CY5 (all from Eurogentec Ltd, Fawley, Hampshire, UK) were used as fluorochrome reporters for the hydrolysis probes analysed in multiplexed reactions with between 2 and 4 genes per run including the control. Cycling parameters were 2 min at 95°C, then 45 cycles of 10 s at 95°C and 60 s at 60°C. Data were collected and analyzed by Rotor-Gene Q Series Software. Gene target Ct (cycle threshold) values were normalized to a Hypoxanthine-Guanine phosphoribosyltransferase 1 (HPRT1) internal control. Ct values were converted to transcript quantity using standard curves obtained by serial dilution of PCR-amplified DNA fragments of each gene. The linear dynamic range of the standard curves covering six orders of magnitude (serial dilution from 3.2 x 10<sup>-4</sup> to 8.2 x 10<sup>-10</sup>) gave PCR efficiencies between 93% and 103% for each gene with R2 &gt; 0.998. Relative gene expression levels after irradiation were determined relative to unexposed controls.</p>

[STEPS]
?. 
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?. 
?. 
?. 
?.
TITLE: No Title
AUTHORS: 

[STEPS]
SECTION: Day Prior to Assay
?. 
SECTION: Materials to Prepare
?. 
SECTION: Procedure
?. 
SECTION: Procedure
?. 
SECTION: Procedure
?. 
SECTION: Procedure
?. 
SECTION: Procedure
?. 
SECTION: Procedure
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SECTION: Procedure
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SECTION: Procedure
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SECTION: Procedure
?. 
SECTION: Procedure
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SECTION: Procedure
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SECTION: Procedure
?. 
SECTION: Procedure
?. 
SECTION: Data Analysis
?.
TITLE: Human Follicular Fluid Procurement and Processing
AUTHORS: hannah.anvari, asma.giornazi, shriya.shah, maryellen.pavone, francesca.e.duncan francesca.duncan

[DESCRIPTION]
Purpose: This protocol is intended for use in the collection and storage of human follicular fluid in a research setting. The protocol details the collection, processing, and long-term storage of follicular fluid for downstream analysis.

[GUIDELINES]
Researchers working with human specimens will adhere to all safety and training protocols required by the institution (Northwestern Medicine/Northwestern University) including but not limited to:    

Biosafety Certification
Bloodborne Pathogens Certification
Collaborative Institutional Training Initiative (CITI program) certification




[STEPS]
SECTION: Collection and processing fluids for storage
1. Follicular fluid is collected from an individual undergoing an egg retrieval procedure in
50 mL conical tubes at  the Fertility and Reproductive Medicine clinic within Northwestern Medicine and stored at 2-4 °C immediately following collection. Research coordinator will collect and bring all tubes of follicular fluid to the lab kept on ice within 1 hour of an egg retrieval procedure. Samples should be transported in blue, labeled biohazard cooler bag (Fig. 1A, B).
   

SECTION: Collection and processing fluids for storage
2. Images are taken of follicular fluid before (Fig. 2A) and after (Fig. 2B) centrifugation and color of samples are noted. Samples will range from straw-colored, which is most ideal, to having a red tint which may indicate blood contamination from the process of egg retrieval and samples can still be stored in this case. Samples are of the pooled follicular fluid in all antral follicles from one patient ovary.  



SECTION: Collection and processing fluids for storage
3. Follicular fluid is centrifuged at 300g for 20min at 4 °C and then the top, clear layer (supernatant) is immediately aliquoted into 2 mL and 5 mL cryovials which are subsequently frozen and stored at -80°C.
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
This protocol describes how to prepare a 100x M2
1x M2 media could be created from the stock and supplemental stocks and trace metals could be added afterwards.

 
Recipes for standard and alternative M2 for culturing freshwater cyanobacteria, such as Synechocystis sp. PCC 6803, in a co-culture togther with heterotrophic bacteria.
Media is usually not suitable for marine cyanobacteria.

Slighly modified from: 
Weiss, Taylor L., Eric J. Young, and Daniel C. Ducat. "A synthetic,  light-driven consortium of cyanobacteria and heterotrophic bacteria  enables stable polyhydroxybutyrate production." Metabolic engineering 44 (2017): 236-245.

[STEPS]
SECTION: Important information
?. 
SECTION: Addition of ingredients for M2 in a 1 l bottle
?. 
SECTION: Addition of ingredients for M2 in a 1 l bottle
?. 
SECTION: Addition of ingredients for M2 in a 1 l bottle
?. 
SECTION: Addition of ingredients for M2 in a 1 l bottle
?. 
SECTION: Addition of ingredients for M2 in a 1 l bottle
?. 
SECTION: Addition of ingredients for M2 in a 1 l bottle
?. 
SECTION: Addition of ingredients for M2 in a 1 l bottle
?. 
SECTION: Addition of ingredients for M2 in a 1 l bottle
?.
TITLE: Contaminated Liquid Disposal
AUTHORS: jack.hocking

[DESCRIPTION]
This protocol is for the cleaning of contaminated glassware in our lab.

[BEFORE_START]
Virkon can be corrosive to lower-quality metals, where possible minimize contact with metal surfaces.


[GUIDELINES]
Virkon should only be used at the concentration specified on the packaging.
Ensure sufficient PPE is worn in line with the Virkon SDS.
Virkon should be disposed of after the pink colour fades, or after 1 week.

[STEPS]
SECTION: Erlenmeyer Flasks
2. Fill flask with Virkon solution to about 3/4 of the total volume and leave covered.
SECTION: Bottles
3. If the bottle is less than half full, fill the remainder with Virkon.
If the bottle is more than half full, pour into a larger container and refill the smaller bottle with Virkon.
If the bottle contains 500 mL or 1000 mL of liquid, deposite an appropriate number of Virkon tablets directly into the bottle.
SECTION: Bottles
4. After adding Virkon to contaminated glassware, leave for 24-48 hours.
SECTION: In-Lab Cleaning
1. Fully dissolve one Virkon tablet per 500 mL of required solution.

SECTION: Bottles
5. Dispose of the Virkon down the sink and send glassware to autoclave.
SECTION: Autoclaving Steps
6. ​All glassware containing contaminated fluid is placed in metal pails, with the exception of our flasks which are left in the tube rack on the autoclave shelf. 

SECTION: Autoclaving Steps
7. ​Two indicators are placed in, one on each shelf, which change colour when 121°C. 
SECTION: Autoclaving Steps
8. A probe is placed in 1 L of water to ensure liquid temperature is reached.
SECTION: Autoclaving Steps
9. The autoclave is run on a No. 3 cycle, 121°C for 60 minutes.
SECTION: Autoclaving Steps
10. The now-sterile liquid is emptied down the sink.
SECTION: Autoclaving Steps
11. Bottles and flasks are soaked in Decon and Chlorus for one hour, scrubbed, and then left to soak for another hour.  Reactor flasks are only soaked in Decon.
SECTION: Autoclaving Steps
12. Glassware is rinsed with deionised water and left to dry in ovens.
SECTION: Autoclaving Steps
13. Once dry, the glassware is moved to the sterilising autoclave which runs at 121°C for 15 minutes.
SECTION: Autoclaving Steps
14. Glassware is left to dry overnight in the sterilising autoclave.
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
This protocol outlines the analysis used to generate input files for CoNet for the phage-bacteria network. Based on methods from the following publication:<br /><br />Hannigan, Geoffrey D., et al. "The Human Skin Double-Stranded DNA Virome: Topographical and Temporal Diversity, Genetic Enrichment, and Dynamic Associations with the Host Microbiome." <em>mBio</em> 6.5 (2015): e01578-15.

[BEFORE_START]
Supplemental information available at:<br /><br /><a href="https://figshare.com/articles/The_Human_Skin_dsDNA_Virome_Topographical_and_Temporal_Diversity_Genetic_Enrichment_and_Dynamic_Associations_with_the_Host_Microbiome/1281248" target="_blank">https://figshare.com/articles/The_Human_Skin_dsDNA_Virome_Topographical_and_Temporal_Diversity_Genetic_Enrichment_and_Dynamic_Associations_with_the_Host_Microbiome/1281248</a>

[GUIDELINES]
<span class="identifier">sessionInfo</span><span class="paren">(</span><span class="paren">)<br /><br />## R version 3.2.0 (2015-04-16) <br />## Platform: x86_64-apple-darwin13.4.0 (64-bit) <br />## Running under: OS X 10.10.4 (Yosemite) <br />## ## locale: <br />## [1] en_US.UTF-8/en_US.UTF-8/en_US.UTF-8/C/en_US.UTF-8/en_US.UTF-8 <br />## <br />## attached base packages: <br />## [1] stats graphics grDevices utils datasets methods base <br />## <br />## loaded via a namespace (and not attached): <br />## [1] magrittr_1.5    formatR_1.2    tools_3.2.0    htmltools_0.2.6 <br />## [5] yaml_2.1.13    stringi_0.4-1    rmarkdown_0.7    knitr_1.10.5 <br />## [9] stringr_1.0.0    digest_0.6.8    evaluate_0.7</span>

[STEPS]
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?. 
?.
TITLE: sCD40L ELISA Assay
AUTHORS: fzhou

[DESCRIPTION]
This protocol for the sCD40L ELISA kit (Cat#BMS6010, Invitrogen) is used to measure the concentration of sCD40L in serum and CSF samples in mice.

[STEPS]
1. Prepare Wash Buffer.
2. Wash the microwell strips twice with 300 μL Wash Buffer per well. Allow the Wash buffer to sit for 10-15s before harshly drop it in the strain. Wash again.
3. Empty wells on the paper towel, use the strip immediately after washing.
4. Prepare the standard dilution as the protocol. Using Sample Diluent (provided) to dilute samples.
5. Add 100 μl of Sample Diluent in duplicate to the blank wells.
6. For serum samples, add 50 μl of Sample Diluent to the sample wells and then add 50 μl of serum samples to the sample well. For CSF samples, add 5 μl of Sample Diluent to the sample wells and then add 95 μl of serum samples to the sample well.
7. Prepare Bio-Conjugate (fresh) and add 50 μl of Bio-Conjugate to all wells.
8. Apply an adhesive film to the plate and let it incubate at room temperature (RT) for 2 hrs on a microplate shaker.
9. Remove adhesive film and empty wells. Wash 4 times using Wash Buffer.
10. Prepare Streptavidin-HRP and add 100 μl of diluted Streptavidin-HRP to all wells.
11. Apply an adhesive film to the plate and let it incubate at RT for 1 hr on a microplate shaker.
12. Remove adhesive film and empty wells. Wash 4 times using Wash Buffer.
13. Add 100 μl TMB substrate solution to all wells. Incubate the strips at RT until the color changes. Avoid light.
14. Add 100 μl the stop solution to all wells.
15. Measure the absorbance of each microwell on a spectro-photometer at a wavelength of 450 nm.