eamag/protocols-with-steps huggingface dataset:
TITLE: Construction of a Moore Swab
AUTHORS: Dilip  Abraham, Nicola Elviss, Nicholas Feasey, Nick  Grassly, Jacob John, Gagandeep Kang, John  Scott Meschke, Venkata  Raghava Mohan, Satheesh Nair, Jonathan  Rigby, Rajan Srinivasan, Catherine  Troman, Christopher  Uzzell, Nicolette Zhou

[DESCRIPTION]
Moore swabs are gauze swabs used to capture bacteria in water such as sewage and wastewater. The gauze is held together by a string which is then tied to a stationary object in the water and left for 24-72 hours. During this time, as water moves past the swab bacteria are caught and attach to the swab, allowing isolation and identification for monitoring and surveillance of the bacteria of interest. This protocol based on the Moore swab construction described in Sikorski and Levine (2020).

[GUIDELINES]
The material used to tie the swab together may depend on the nature of the water it is being placed in. For example, if it is faster flowing or there are many solids in the water then you may require a stronger fishing line rather than twine to reduce loss of swabs. 

[STEPS]
1. Fold the gauze eight times length-wise to form an 8-ply square
2. Tie the gauze around the middle with twine/fishing line leaving a long tail for attaching the swab once it has been placed.
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
In a study comparing obese versus lean twins, Turnbaugh and colleagues found that there were "shared microbial genes among sampled individuals, comprising an extensive, identifiable 'core microbiome' at the gene, rather than at the organismal lineage level".  <br />This means that despite having differences in bacterial composition, the "core" functional repertoire of microbiome remains the same in obese versus lean twins.  <br />In this tutorial, we are going to explore these functional differences in two lean twins (TS1 and TS2) and two obese twins (TS19 and TS20) from this study.   <br /><br />Turnbaugh, Peter J., et al. "A core gut microbiome in obese and lean twins."<em>nature</em> 457.7228 (2009): 480-484.

[GUIDELINES]
Read the paper here:<br />http://www.nature.com/nature/journal/v457/n7228/abs/nature07540.html

[STEPS]
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TITLE: Exp01_protocolo
AUTHORS: Julio Cesar Quintero Gámez

[DESCRIPTION]
Nuestro experimento consistió en investigar el comportamiento en la temperatura de una taza a 68 °C expuesta a una temperatura ambiente de 19 °C. Utilizamos sensores DS18B20 para medir tanto la temperatura del agua como la del entorno.  Los resultado nos mostraron la comprobación de la ley 0 ya que los cuerpos terminan con la misma temperatura con el paso del tiempo, además se observo una peculiaridad, la disminución de la temperatura no fue lineal, sino que describió una trayectoria exponencial.

[GUIDELINES]
Asegúrese de mantener el experimento en un ambiente con la temperatura lo mas controlada posible.

[STEPS]
SECTION: Código y Arduino
1. Instalación del software a utilizar.
SECTION: Código y Arduino
2. Armado del circuito

 

SECTION: Experimento
3. Vierte 100 mL de agua en tu olla. Enciende la parrilla y toma mediciones cada cierto tiempo para saber cuando el agua llegue a 68 °C para verterlo a la taza.
SECTION: Experimento
4. Introduce uno de los sensores en el agua y el otro déjalo expuesto al ambiente.
SECTION: Experimento
5. Dejar el experimento corriendo hasta que se observe que ah llegado al equilibrio térmico, esto puede durar un par de horas.
SECTION: Experimento
6. Extraer los datos obtenidos.
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p>Experiment purpose: measure dynamics of phage infection and gene expression in cyanobacteria. We collect samples every 1-2 hours over a 12 hour period, starting from an initial infection volume of 200ml per replicate per treatment.</p>

[STEPS]
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TITLE: Making tetracycline LB agar plates
AUTHORS: Bonnie Evans

[DESCRIPTION]
Making LB agar plates containing tetracycline for selecting strains carrying the pJC8 cosmid. Plates can be stored at 4 °C wrapped in aluminium foil for up to a month. 

[STEPS]
SECTION: Making tetracycline LB agar plates
1. Autoclave or microwave LB agar from media room

 

SECTION: Making tetracycline LB agar plates
2. Make up 15 mg/ml tetracycline hydrochloride in sterile water
SECTION: Making tetracycline LB agar plates
3. Let LB agar cool to 55 °C
SECTION: Making tetracycline LB agar plates
4. Dilute tetracycline hydrochloride 1/1000 dilution in LB agar (final conc: 15 ug/mL).
SECTION: Making tetracycline LB agar plates
5. Using a serological pipette, dispense 35 mL agar into 90 mm Petri dishes. Leave to set. 


SECTION: Making tetracycline LB agar plates
2.1. Retrieve tetracycline hydrochloride from -20 °C freezer
SECTION: Making tetracycline LB agar plates
2.2. Weigh an empty Eppendorf
SECTION: Making tetracycline LB agar plates
2.3. In the fume hood, add tetracycline hydrochloride to the Eppendorf
SECTION: Making tetracycline LB agar plates
2.4. Weigh the Eppendorf, and calculate the difference to determine the weight (mg) of compound
SECTION: Making tetracycline LB agar plates
2.5. To calculate sterile water (uL) to add =  (compound mg / 15 mg/mL) x 1000 uL
TITLE: Retina and RPE/Choroid RNA Extraction Protocol
AUTHORS: Chottiwatt Jittprasong

[DESCRIPTION]
This protocol involves the extraction of RNA from retinal and retinal pigment epithelium (RPE) cells in the subject's retina, utilizing TRIzol as the main reagent for cell lysis.

[BEFORE_START]
Prior to the initiation of this protocol, extraction of the retinal sample from the subject's eyeball is required.

[STEPS]
SECTION: RNA Extraction
1. Precool the centrifuge to 4 °C 

SECTION: RNA Extraction
2. Add 1000 µL  of TRIzol per retina and pipette up and down 30 times to homogenize the mixture.
 

SECTION: RNA Extraction
3. Incubate for 5 min on ice 

SECTION: RNA Extraction
4. Add 200 µL of Chloroform per 1000 µL of TRIzol used for lysis and mix thoroughly.

SECTION: RNA Extraction
5. Incubate for 3 min on ice 

SECTION: RNA Extraction
6. Centrifuge the sample at 12000 x g, 15 min, 4 °C 

SECTION: RNA Extraction
7. Transfer the aqueous phase containing the RNA to a new tube. 
 

SECTION: RNA Extraction
8. Add 500 µL of Isopropanol to the aqueous phase, per 1000 µL of TRIzol used.

SECTION: RNA Extraction
9. Add 4 µL of Glycoblue and mix well.

SECTION: RNA Extraction
10. Incubate at -4 °C for 60 min 

SECTION: RNA Extraction
11. Centrifuge at 30000 x g, 15 min, 4 °C 
 

SECTION: RNA Extraction
12. Discard supernatant carefully.
SECTION: RNA Extraction
13. Resuspend the pellet in 1000 µL of on ice 75% ethanol per 1000 µL of TRIzol. Vortex and centrifuge at 7500 x g, 5 min, 4 °C 
SECTION: RNA Extraction
14. Repeat Step 13.
SECTION: RNA Extraction
15. Air dry the RNA pellet for 5 min 

SECTION: RNA Extraction
16. Resuspend the pellet in 20-50 µL of RNase free water.
 


SECTION: RNA Extraction
17. Quantify the sample by NanoDrop
TITLE: Assessing Astrocyte Territory Volume and 3D Sholl Analysis
AUTHORS: Shiyi Wang

[DESCRIPTION]
Assessing Astrocyte Territory Volume and 3D Sholl Analysis

[STEPS]
1. **Section Collection**
1.1. - Collect 100 μm-thick floating sections containing the anterior cingulate cortex (ACC) and primary motor cortex (MOp) from mice.
1.2. - Ensure astrocytes are sparsely labeled via PALE with mCherry-CAAX.
2. **Imaging**
2.1. - Use an Olympus FV 3000 microscope with a 60x objective to acquire high-magnification images (50-60 μm z-stack) of individual astrocytes.
2.2. - Criteria for inclusion: Capture the entire astrocyte within a single brain section, located in layer 2/3 of ACC or MOp.
3. **Surface Reconstructions**
3.1. - Analyze imaged astrocytes using Imaris Bitplane software.
3.2. - Generate surface reconstructions of the astrocytes.
4. **Territory Volume Measurement**
4.1. - Use Imaris Xtensions “Visualize Surface Spots” and “Convex Hull” to create additional surface renders representing the territory of each astrocyte.
4.2. - Record the volume of each astrocyte territory.
5. **Statistical Testing**
5.1. - Analyze astrocyte territory sizes from biological replicates across experimental conditions.
5.2. - Use a nested two-way ANOVA followed by the Bonferroni posthoc test for statistical comparisons.
6. **Surface and Filament Creation**
6.1. - Load astrocyte images onto Imaris and create a surface representation for each astrocyte.
7. **Filament Generation**
7.1. - Create filaments using ‘Add new filament (leaf icon)’ in Imaris.
8. **Quantification of Complexity**
8.1. - Use the gear tool on Imaris to display Sholl intersections for each astrocyte.
8.2. - Quantify the complexity of astrocytes based on Sholl intersections.
9. #### Experimental Details
10. ### Notes:
11. - Maintain consistency in imaging and analysis methodologies across experimental conditions.
12. - Consider biological variability and replicate experiments to ensure robust conclusions.
TITLE: No Title
AUTHORS: 

[STEPS]
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TITLE: Hemocytometer use to quantify concentracion of cells&#39; suspensions
AUTHORS: abotero  

[DESCRIPTION]
This protocol presents the procedures to estimate the concentration of cells' suspensions using an hemocytometer

[GUIDELINES]
The lower limit for accurate counting of cells in a hemocytometer is usually considered to be 2.5 x 105/ml. 

For cell numbers greater than 2.5 x 106/ml, it is generally recommended that the sample be diluted.

[STEPS]
1. Wash the hemocytometer with a washer bottle and dry it using a paper towel
2. Cover the hemocytometer with the coverslip and place it on a flat surface 
3. Vortex briefly the cell suspension whose concentration will be calculated
4. Using a micropipette collect 10-100 µl of the previously agitated suspension
5. Place the micropipette tip on the hemocytometer edge near the coverslip
6. Empty the spore suspension slowly into the chamber of the hemocytometer
7. Place the hemocytometer in the microscope's mechanical stage and secure it using the stage clip
8. Turn on the microscope and focus on the hemocytometer
9. Look out for the first square to be counted
10. Count the number of cells on each square
 

11. Count all the squares following a zig-zag 
 

12. Write down the number of cells on each square
 

 

13. Estimation of the concentration
 

13.1. If cells were counted on squares of size 1

Concentration (cells/ml)= Number of counted cells*10,000/Number of squares counted

 

13.2. If cells were counted on squares of size 2

Concentration (cells/ml)= Number of counted cells*160,000/Number of squares counted
 

13.3. If cells were counted on squares of size 3

Concentration (cells/ml)= Number of counted cells*250,000/Number of squares counted
TITLE: General fungi ITS2 (ITS4ngsUni - fITS7) for Illumina amplicon sequencing 
AUTHORS: Roey Angel, Eva Petrova

[DESCRIPTION]
A general assay for the preparation of PCR amplicons for further analysis of fungal communities by Illumina amplicon sequencing.

The primers target the 2nd internal transcribed spacer region (ITS2) of the ribosomal RNA gene and were specifically designed for Illumina amplicon sequencing. The ITS4ngsUni primer was designed by Tedersoo et al. (2016) as a universal primer for nearly all eukaryotes, and the fITS7 primer was designed by Ihrmark et al. (2012) as a fungal-specific primer. The ITS2 region primer pair should capture a higher diversity compared to primers targeting the ITS1 region. 

For barcoding, we use the Fludigm Access Array, and therefore, the primers are synthesized with the CS1 and CS2 regions.

When working with soil samples, we usually obtain amplicons in the range of 300-500bp. Double bands are often observed.

[STEPS]
SECTION: PCR reaction
2. Prepare the following master mixture  on ice .
Do not forget to prepare some additional mixture for the negative (NTC = no template) and positive controls, and to account for pipetting errors. Work in a clean PCR box.
 

 
  
  
  


SECTION: PCR program
5. Run the following PCR program:
1.  95 °C 10 min
2.  x 30{
       a.  95 °C 45 s
       b. 52 °C 45 s
       c.  72 °C45 s
      }
4.  72 °C 10 min
5. 10 °C   

SECTION: Primers
1. 
 

SECTION: PCR reaction
3. Vortex and spin down 2 min .

SECTION: PCR reaction
4. Distribute 24 µL  of the mixture to each tube or well of 96-well plate and add 1 µL of template DNA or cDNA.

SECTION: Evaluate PCR products on an agarose gel
6. Prepare a 1.5% agarose gel by mixing:
 
100 mL TAE  
1.5 g agarose  
Heat in the microwave until dissolved and pour into a gel frame.
Place solid gel into an electrophoresis bath filled with TAE buffer.

 
 
 

SECTION: Evaluate PCR products on an agarose gel
7. Load 5 µL of the sample into a well.
In addition load 5 µL of DNA ladder mix (80-10,000 bp) into an empty well, as a marker.

SECTION: Evaluate PCR products on an agarose gel
8. Run the gel at 110 V, 265 mA for approx.40 min

SECTION: Evaluate PCR products on an agarose gel
9. Stain gel for at least 30 min in an Ethidium bromide TAE bath (or any other DNA stain). 
 


SECTION: Evaluate PCR products on an agarose gel
10. Visualize the gel using a gel documentation system.
TITLE: READDI protocol: Crystallisation of CHIKV nsP3 macrodomain
AUTHORS: Jasmin Aschenbrenner, Peter Marples, michael fairhead, Andre Schutzer de Godoy, Daren Fearon

[DESCRIPTION]
The crystallization protocol and buffer conditions used to obtain reproducible Chikungunya Virus NS3 macrodomain crystals suitable for XChem fragment screening.

[GUIDELINES]
N/A

[STEPS]
SECTION: Crystallisation experiment
3. Protein and buffer requirements:
21.6 µL11 mg/mL  
2.88 mL 
10.08 µL   Seeds, dilution 1:100

SECTION: Crystallisation experiment
5. Dispense 30 µL into SwissCI 3 lens plate reservoir wells using a 100 µl multi-channel pipette.
Dispense 75 nL11 mg/mL    to each lens using the SPT mosquito.
Dispense  40 nL to each lens using the SPT mosquito.
Dispense  35 nL to each lens using the SPT mosquito.

Drop ratio: 15:8:7 ratio (75 nl   : 40 nl Crystallisation solution: 35 nl Seeds)
Final drop volume: 150 nl
SECTION: Crystallisation experiment
4. Crystallisation screen composition:
0.1 Molarity (M)  Tris-NaOH pH 7.8  
0.1 Molarity (M)  Potassium thiocyanate
0.1 Molarity (M) Sodium bromide
25 % v/v PEG Smear Broad

Stock solutions used:
1 Molarity (M) Tris adjusted to pH 7.8 with NaOH
1 Molarity (M)  Potassium thiocyanate
1 Molarity (M)  Sodium bromide
50% v/v PEG Smear Broad

 

SECTION: Crystallisation experiment
7. 
 


SECTION: Crystallisation experiment
6. Incubate at 20 °C for 1440 min in Formulatrix Rock Imager.

Imaging Schedule: The first images are taken after 12 h and the imaging schedule follows a Fibonacci sequence of days for further collections. 
SECTION: Equipment needed
1. Formulatrix Rock Imager (or incubator of choice)
SPT mosquito 
 
P100 8 multi-channel pipette
SwissCI 3 lens plate

SECTION: Crystallisation experiment
2. Prepare seed stock:
 
1: 100 dilution    seeds
SECTION: Data collection at Synchrotron
8. Diamond Light Source
Unattended Data Collection (UDC)
Data Collection Temperature: 100K
Detector: DECTRIS EIGER2 X 9M
Beamline: I04-1
Wavelength:  0.9212 Å
Resolution (Å): 1.64
Beam Size (μm): 60 X 50
Number of images: 3600
Oscillation: 0.10°
Exposure (s): 0.0020
Transmission (%): 100
Flux (ph/s): 3.80e+12
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p><em>Asterionella formosa</em> is a freshwater diatom and as such as a silica cell wall. Therefore classical protocols for RNA extraction do no work.  We have settled this protocol that gives high quality RNA (tested by NGS). This protocol was also applied to marine diatoms, <em>Thalassiosira pseudonana</em> and <em>Phaeodactylum tricornutum</em> and gave very good yields.</p>

[STEPS]
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TITLE: No Title
AUTHORS: 

[STEPS]
SECTION: Reaction Setup on ice
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SECTION: Reaction Setup on ice
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SECTION: Reaction Setup on ice
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SECTION: Reaction Setup on ice
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SECTION: Reaction Setup on ice
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SECTION: Reaction Setup on ice
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SECTION: Reaction Setup on ice
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SECTION: Thermocycling conditions
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SECTION: Thermocycling conditions
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SECTION: Thermocycling conditions
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SECTION: Thermocycling conditions
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TITLE: Determining Genome Targeting Efficiency using T7 Endonuclease I (M0302)
AUTHORS: New  England   Biolabs

[DESCRIPTION]
T7 Endonuclease I recognizes and cleaves non-perfectly matched DNA. This protocol describes how to determine genome targeting efficiency by digesting annealed PCR products with T7 Endonuclease I. In the first step PCR products are produced from the genomic DNA of cells whose genomes were targeted using Cas9, TALEN, ZFN etc. In the second step, the PCR products are annealed and digested with T7 Endonuclease I. Fragments are analyzed to determine the efficiency of genome targeting.

[GUIDELINES]
REFERENCES:
 


[STEPS]
SECTION: PCR
1. Set up a 50 µL PCR reaction using ~100 ng as a template. For each amplicon set up 3 PCR reactions using the following templates:
gDNA from targeted cells (e.g. Cas9, or TALEN transfected cells)
gDNA from negative control cells (e.g. non-specific DNA transfected cells)
water (i.e. no template control)
PCR using Q5 High-Fidelity DNA Polymerase
   Component 50 µl reaction Final Concentration   Q5® Hot Start High-Fidelity 2X Master Mix (M0494S) 25 µl 1X   10 µM Forward Primer 2.5 µl 0.5 µM   10 µM Reverse Primer 2.5 µl 0.5 µM   Template DNA variable 100 ng total   Nuclease-free water To 50 µl     

SECTION: PCR
2. Gently mix the reaction.
SECTION: PCR
3. Collect all liquid to the bottom of the tube by a quick spin if necessary.
SECTION: PCR
4. Transfer PCR tubes to a PCR machine and begin thermocycling:
Cycling Conditions
 

 
SECTION: PCR
5. Analyze a small amount of the of the PCR product to verify size and appropriate amplification.
SECTION: PCR
6. Purify the PCR reaction using 90 µL following the manufacturer’s recommendations.  
 

SECTION: PCR
7. Elute PCR products in 30 µL, recovering 25 µL.
SECTION: PCR
8. Measure the concentration of the purified PCR products.
SECTION: T7 Endonuclease I digestion
9. Assemble reactions as follows:  
 

SECTION: T7 Endonuclease I digestion
10. Anneal the PCR products in a thermocycler using the following conditions:
Hybridization Conditions
 

SECTION: T7 Endonuclease I digestion
11. Add 1 1 to the annealed PCR products for a final volume of 20 µL. 
SECTION: T7 Endonuclease I digestion
12. Incubate at 37 °C for 15 min.
SECTION: T7 Endonuclease I digestion
13. Stop the reaction by adding 1.5 µL. 
SECTION: T7 Endonuclease I digestion
14. Purify the reaction using 36 µL according to the manufacturer’s suggestion.   
 

SECTION: T7 Endonuclease I digestion
15. Elute the DNA fragments in 20 µL, recovering 15 µL.
SECTION: Analysis
16. Analyze the fragmented PCR products and determine the percent of nuclease-specific cleavage products (fraction cleaved).
SECTION: Analysis
17. Calculate the estimated gene modification using the following formula:
% gene modification = 100 x (1 – (1- fraction cleaved)1/2)
TITLE: Staphylococcal Protein-A and Chimeric Protein-LAG sandwich ELISA
AUTHORS: Angel  Justiz-Vaillant

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">This ELISA was used to study the interactions between Staphylococcal protein-A (SpA) and protein-LAG (PLAG) with different immunoglobulin preparations from mammalian and avian species. </div></div>

[STEPS]
?. This ELISA was used to study the interactions between Staphylococcal protein-A (SpA) and protein-LAG (PLAG) with different immunoglobulin preparations from mammalian and avian species. The 96 well microtiter plate was coated overnight at 4°C with 2 µg/µl per well of SpA in carbonate-bicarbonate buffer pH 9.6.
?. The plate was then treated with bovine serum albumin solution and washed 4X with PBS-Tween. 50 µl of immunoglobulins (1 mg/ml) is added and incubated for 1h at room temperature, and the microplate is rewashed 4X with PBS-Tween.
?. Then, 50 µL of peroxidase-labeled PLAG conjugate diluted 1:5000 in PBS-non-fat milk was added to each well and incubated for 1h at RT. The plate was washed 4X with PBS-Tween.
?. Then, 50 µL of o-phenylenediamine solution (4 mg/mL) was added, and the plate was incubated for 15 min at RT in the dark. The reaction was stopped with 50 µL of a 3M H2SO4 solution.
?. The plate was visually assessed for color development and read on a microplate reader at 492 nm. A cut-off point was calculated as the mean of the optical density of the negative controls multiplied by two. The cut-off value was set to 0.32.
TITLE: 10x Protocols: Visium v1 FFPE Library Construction -- University of Minnesota TMCs (CG000407 Rev D)
AUTHORS: 10x Genomics, Laura J Niedernhofer , David A Bernlohr

[DESCRIPTION]
Protocols from 10x Genomics for Visium Spatial Gene Expression v1 chemistry on FPPE samples (without the CytAssist component).

1. Protocol ID# (CG) and Revision used: 
    10x protocol CG000407, Revision D -- Library Construction 

2. Additional protocols added for guidance:
    10x protocol for imaging guidelines 
    10x protocol for tissue preparation guidelines 

Note: These protocols may not be the current version offered by 10x Genomics but were used to produce the specific datasets connected to them. Please review the 10x Genomics support website for the most recent versions of the protocols prior to starting your experiment. 

[STEPS]
1. 10x protocol CG000407, Revision D (Library construction)
  

2. Additional Protocols/Resources
  
  
https://www.10xgenomics.com/support/spatial-gene-expression-ffpe
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p>Glutamate is enzymatically measured in the cerebrospinal fluid by monitoring the fluorescence increase due to NADPH<sup>+ </sup>production in the presence of glutamate dehydrogenase and NADP<sup>+ </sup>on a spectrofluorimeter (Shimadzu RF-5301PC, Japan).</p>

[STEPS]
TITLE: N_terminal protein labeling
AUTHORS: Patricia Yuste Checa, F Ulrich Hartl

[DESCRIPTION]
This protocol details how to efficiently label a protein at the N-terminus using Clusterin protein as example. 

[STEPS]
SECTION: N-terminal protein labeling
1. Exchange the protein buffer to Labeling buffer using a Nap-5 column (Thermo Fisher Scientific, 45-000-151). Equilibrate the column with 10 column volumes (CV: 1 mL). Load the protein onto the column and elute the protein with the corresponding amount of Labeling buffer following the column manufacturer’s instructions. Collect each eluted drop into an Eppendorf low binding tube. Measure the protein concentration in each fraction by nanodrop and pool the protein-containing fractions.
 

SECTION: N-terminal protein labeling
2. Dissolve the dye (Alexa488 NHS ester(refer materials section); pHrodo Red Succinimidylester in DMSO. With a pipette tip gently touch the dye powder which will stick to the tip. Immerse the tip in some DMSO previously dispensed in a tube. Repeat the procedure several times until the solution reaches the desired color.
 

SECTION: N-terminal protein labeling
3. Quantify the diluted dye concentration by nanodrop. Dilute the dye with water for measurement in order to reach an absorbance of λ<1 for an accurate measurement.
 


SECTION: N-terminal protein labeling
4. Add the corresponding amount of diluted dye to the eluted protein to reach a final protein:dye ratio of 1:4-1:10.
SECTION: N-terminal protein labeling
5. Incubate 90 min at Room temperature in the dark.

SECTION: N-terminal protein labeling
6. Remove free dye by using a Nap-5 column, pre-equilibrated with 1x PBS pH 7.2 buffer or desired final buffer. Collect each eluted drop in an Eppendorf low binding tube. Measure protein concentration in each fraction by nanodrop and pool the fractions containing protein.
 


SECTION: N-terminal protein labeling
7. Measure labeling efficiency (dye molarity/protein molarity) by nanodrop. Quantify independently dye concentration and protein concentration.
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
For use in <a href="https://www.protocols.io/view/DNase-I-Treatment-c3myk5" target="_blank">DNase I Treatment</a> protocol for DNase inactivation

[GUIDELINES]
Note: EGTA tetratsodium salt is far easier to get into solution than the disodium salt. It is possible to get to 1.5M with this reagent.

[STEPS]
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