eamag/protocols-with-steps huggingface dataset:
TITLE: Competent Transformation
AUTHORS: Miquel Estévez-Gay

[DESCRIPTION]
Transform a Plasmid into previously obtained Competent E.coli Cells. 

[STEPS]
1. Melt in ice and aliquot of 100μL competent E-coli DH5α or BL21(DE3) . 
2. Preheat 500 µL LB or SOC medium (37ºC). 

3. Add 50-100μg DNA into competent cells and mix softly. This is usually 1-2µl from minipreps. 
I recommend using 10µl if the DNA comes from PCR and/or DpnI digestion. 
4. Incubate 20 min in ice. 

5. Incubate without shaking at 42 °C for1 min. 

6. Incubate 2 min in ice. 
7. Add 500 µL LB or SOC pre-heated media. This must be done under Laminar Flux Cabin

8. Incubate 60 min  in shaker thermal block. 

9. Prepare Amp or Kan plates. 
10. Add 100 µLcompetent cells into each plate and plate. Mix using sterile glass balls under laminar flux cabin. 

11. Incubate 37 °C60 min 

12. Next day, store the plates parafilmed in the fridge. Can be stored for several weeks.
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p>This protocol will introduce a workflow for quality control and pre-processing of metagenomic sequence reads using FastQC for visualization and FastX Toolkit for editing the fastq files. </p>

[GUIDELINES]
<p><a href="http://hannonlab.cshl.edu/fastx_toolkit/index.html" target="_blank">FastX Toolkit Documentation</a></p>

[STEPS]
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TITLE: Mechanistic Assays (Part 7 of Safety and Efficacy of Imatinib for Preserving Beta-Cell Function in New-onset Type 1 Diabetes Mellitus)
AUTHORS: Stephen.Gitelman , Jeffrey A. Bluestone

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">This is Part 7 of "Safety and Efficacy of Imatinib for Preserving Beta-Cell Function in New-Onset Type 1 Diabetes Mellitus".</div><div class = "text-block">This clinical study is supported by JDRF. The aim of the collection is to determine whether imatinib will slow the progression of the autoimmune destruction of ß cells and lead to the preservation of C-peptide secretion in T1DM and to assess Diabetes-related objectives and safety of Imatinib in new-onset type 1 diabetes mellitus".</div></div>

[STEPS]
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p style="text-align: center;"><img style="display: block; margin-left: auto; margin-right: auto;" src="https://s3.amazonaws.com/pr-journal/hnnfjtn.png" width="100" height="90" data-src="https://s3.amazonaws.com/pr-journal/hnnfjtn.png" data-ofn="nehirarma-üst-Tr.png" />  </p>
<p style="text-align: center;">Nehir Biyoteknoloji Ltd. <a href="http://www.nehirbt.com.tr/en/" target="_blank">www.nehirbt.com</a></p>

[BEFORE_START]
<p>This protocol is derived from</p>
<p>[Reference: GeneMATRIX Quick Blood DNA Purification Kit, version 1.2,  EURx company<strong>]</strong></p>

[STEPS]
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TITLE: No Title
AUTHORS: 

[DESCRIPTION]
Drosophila Tissue Processing for Paraffin Embedding<br />modified by the Baehrecke Lab from published protocols by Ristifo and White, 1991 (Pubmed ID&nbsp;1936557) and Muro et al., 2006 (Pubmed ID&nbsp;16887831)

[BEFORE_START]
Collect white pre-pupae, raised at 25 degrees Celsius, every forty minutes until desired number is obtained. Age for 24 hrs at 25 degrees Celsius and then begin fixation based on the time collected for each individual.&nbsp;

[STEPS]
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TITLE: TEST: Ultra Turrax Tube Drive
AUTHORS: Nannie 

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Evaluation of tubes for the Ultra-Turrax Tube Drive instrument from IKA</span></div><div class = "text-block">Date: 10th February 2021 + 17th February 2021</div><div class = "text-block">Consumables: 1 petri dish 5 steel beads; </div><div class = "text-block">Tubes: </div><div class = "text-block">10/2</div><div class = "text-block">- plastic rotor stator element ("rs")</div><div class = "text-block">- 10 provided steel beads (5 mm) ("steel10")</div><div class = "text-block">- 10 provided glass beads (6 mm) ("glass")</div><div class = "text-block">- 10 provided steel beads + 5 steel beads from DNA lab (3.2 mm) ("steel10+5")</div><div class = "text-block">- disperser with metal rotator ("disp")</div><div class = "text-block">17/2</div><div class = "text-block">- rs ("II")</div><div class = "text-block">- 10 provided steel beads + 5 steel beads from DNA lab (3.2 mm) + 10 silica beads (2.3 mm) ("10+10+10")</div><div class = "text-block">Test sample:</div><div class = "text-block">SYM37M; L1</div><div class = "text-block">Instrument settings:</div><div class = "text-block">10/2</div><div class = "text-block">6000 rpm</div><div class = "text-block">Homogenization for 30s + 30s</div><div class = "text-block">17/2</div><div class = "text-block">6000 rpm</div><div class = "text-block">Homogenization for a total of 120s</div><div class = "text-block">Evaluation:</div><div class = "text-block">The rs created the smallest pieces, while the beads created the largest pieces. The disp was somewhere in between.</div><div class = "text-block">Future prediction:</div><div class = "text-block">A rotor element in steel would propbably work even better than the plastic rs. The Tube Mill 100 instrument could work well.</div></div>

[STEPS]
?. rs on the instrument (second 30s run).Steel10+5 on the instrument (second 30s run).
TITLE: Assessment of autophagic flux assay in the adult Drosophila brain
AUTHORS: Mel Feany

[DESCRIPTION]
Transgenic flies expressing GFP-mCherry-Atg8a (obtained from the Bloomington Drosophila Stock Center)  
 under the control of the  neuronal nSyb-GAL4 driver are used for assaying autophagic flux.

[STEPS]
1. Transgenic flies expressing GFP-mCherry-Atg8a (obtained from the Bloomington Drosophila Stock Center)  under the control of the  neuronal nSyb-GAL4 driver are used for assaying autophagic flux
2. Brains from 10-day-old animals are dissected in ice-cold 1X PBS
3. The dissected brain are transferred to a glass slide
4. The remaining 1X PBS is removed from the slide
5. A drop of DAPI Fluoromount is added and the brain mounted with a coverslip
6. The dissected brain is imaged immediately within 5 minutes using a Zeiss confocal microscope under a 63X objective lens, and a line scan performed
7. For quantification, the intensity of GFP and mCherry is measured per puncta using ImageJ, and the ratio between GFP and mCherry calculated
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
Generation of DNA fragments by DNase digestion involves digesting DNA for a range of times, then picking the time that gives optimal-sized DNA fragments (typically 1000–4000 bp).

[STEPS]
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TITLE: Goga Lab RT-qPCR protocol: QuantStudio6 Machine
AUTHORS: Jeremy.williams

[DESCRIPTION]
Guidelines for preparing RT-qPCR samples for QuantStudio 6 located in HSW7 lab space.

[BEFORE_START]
*Thaw on ice (leave time!) and keep all reagents on ice through preparation. Prepare the plate on ice.


[GUIDELINES]
*start with full tip boxes and use tips in coordination with your plate map, so you never get lost
*watch 1uL volumes in the pipet tip like a hawk - major source of variability
*cap and gently vortex your *mixture per primer set* every 12 replicates or so, sometimes things separate

[STEPS]
SECTION: Prior to plate preparation:
1. Dilute stock IDT primers to 100uM (see note in Guidelines).
2. Use 'GogaLab-RTqPCR-Preparation' Excel spreadsheet to input your sample number and calculate reagent volumes.

SECTION: Prepare your plate:
3. Mix *total reagent volumes required* (see spreadsheet, green) for DI and PowerUP
SECTION: Prepare your plate:
4. Mix your forward and reverse primer pairs together, to a final dilution of 10uM forward and 10uM reverse. For example, add 10uL each of forward and reverse primers to 80uL PCR-quality DI for 100uL final volume.
SECTION: Prepare your plate:
5. Mix *mixtures per primer set* (blue) volumes together.
SECTION: Prepare your plate:
6. Pipet 1uL diluted cDNA into respective qPCR well, aiming for the sidewall of each respective well.
SECTION: Prepare your plate:
7. Add 19uL *mixture per primer set* into each respective well.
SECTION: Prepare your plate:
8. Seal plate, and spin down 1000rpm for 1 minute. Use bacterial, not tissue culture, centrifuge.   
SECTION: Prepare your plate:
9. Load plate into centrifuge and proceed using QuantStudio software suite.
TITLE: Live-cell imaging
AUTHORS: michela.deleidi, María José Pérez J.

[DESCRIPTION]
Live-cell imaging is a technique to visualize dynamic cellular processes in living biological samples.

[STEPS]
SECTION: 1. Cell Preparation
1. Wash cells 1x with OptiMEM (Gibco).

SECTION: 1. Cell Preparation
2. Incubate cells with 100nM MitoTracker red CMH2Xros (Thermo Fisher Scientific) in OptiMEM at 37 °C for 30 min
SECTION: 1. Cell Preparation
3. Wash cells with OptiMEM once, and keep cells in the same medium for imaging
SECTION: 2. Imaging
4. Images were acquired using a Leica TCS SP8 confocal microscope (Leica, Germany) with a 100×/1.4 numerical aperture oil-immersion objective.
SECTION: Procedure for labelled alpha-synuclein Pre-formed Fibrils Experiments
6. Treat iPSC-derived neurons with 0.25 μM Alexa Fluor 594-labeled PFFs (594-PFF) with or without cotreatment with 1 μM CDDO-Me (Cayman Chemical)
SECTION: Procedure for labelled alpha-synuclein Pre-formed Fibrils Experiments
7. After 24 h, incubate cells with 100 nM MitoTracker Green (Invitrogen, MA, USA) in a neuronal medium for 30 min at 37 °C
SECTION: 2. Imaging
5. Analyze images using Diffraction PSF 3D and DeconvolutionLab2 plugins in Fiji-ImageJ version 2.3.0/1.53q (https://fiji.sc; RRID:SCR_002285)
SECTION: Procedure for labelled alpha-synuclein Pre-formed Fibrils Experiments
8. Use ACellBrite™ Steady 488 Membrane Staining Kit (Biotium) to visualize cell membranes following the manufacturer's instructions
SECTION: Procedure for labelled alpha-synuclein Pre-formed Fibrils Experiments
9. Images were acquired using a Leica TCS SP8 confocal microscope with a 63 × /1.4 numerical aperture oil-immersion objective
SECTION: Procedure for labelled alpha-synuclein Pre-formed Fibrils Experiments
10. Acquire Z-stacks for the calculation of the PFF particle area and fluorescence intensity
SECTION: Procedure for labelled alpha-synuclein Pre-formed Fibrils Experiments
10.1. For each condition, 5-8 images were acquired from at least four independent experiments
SECTION: Image processing
11. For the quantification of colocalization and image processing, images were analyzed using the “Analyze particles” and “EzColocalization” plugins in Fiji-ImageJ
TITLE:     ELISA for quantification of IL-19 in human serum or plasma.
AUTHORS: Angel  Justiz-Vaillant

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Interleukins (IL) are a type of cytokine first thought to be expressed by leukocytes alone but have later been found to be produced by many other body cells. They play essential roles in the activation and differentiation of immune cells, as well as proliferation, maturation, migration, and adhesion. They also have pro-inflammatory and anti-inflammatory properties. The primary function of interleukins is, therefore, to modulate growth, differentiation, and activation during inflammatory and immune responses. Interleukins consist of a large group of proteins that can elicit many reactions in cells and tissues by binding to high-affinity receptors in cell surfaces. </div><div class = "text-block"><span>Th2 lymphocytes synthesize IL-19 and acts on resident vascular cells in addition to immune cells. It is an anti-inflammatory molecule. It promotes immune responses mediated by regulatory lymphocytes and has substantial activity on microvascular.</span><a href="https://www.ncbi.nlm.nih.gov/books/NBK499840" style = "text-decoration:underline;color:blue;cursor:pointer;"><span style = ":;">[1]</span></a></div><div class = "text-block">Reference</div><div class = "text-block"><span>1. Justiz Vaillant AA, Qurie A. Interleukin. In:</span><span style = "font-style:italic;">StatPearls</span><span>. Treasure Island (FL): StatPearls Publishing; June 12, 2019.</span></div></div>

[STEPS]
?. An anti-human IL-19 coating antibody is adsorbed onto microwells by incubation overnight at 4°C.
?. Add 50 µl of human serum. Human IL-19 present in the serum sample binds to antibodies adsorbed to the microwells.
?. The microplate is blocked with 3% non-fat milk-PBS buffer and later wash to remove unbound proteins.
?. Fifty (50) µl of biotin-conjugated anti-human IL-19 antibody is added. The optimal dilution must be investigated.
?. The microplate is rewashed with PBS-Tween 20 buffer, pH 7.4.
?. One hundred µl of streptavidin-HRP conjugate is added and it binds to the biotin-conjugated anti-human IL-19 antibody. The optimal dilution of this conjugate must be investigated.
?. The plate is washed following incubation to remove the unbound Streptavidin-HRP.
?. Add 100 µl of 3',3',5',5'- tetramethylbenzidine (TMB; Sigma-Aldrich) to each well.
?. Incubate the microwells in the dark for 15 min.
?. A colored product is formed in proportion to the quantity of human IL-19 present in the sample or standard.
?. The reaction is terminated by addition of 100 µl 3M H2SO4  and absorbance is measured at 450 nm.
?. A standard curve is made from 7 human IL-19 standard dilutions and the human IL-19 sample concentration is determined.
?. For better results place the microplate on a microplate shaker in every incubation.
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<div>
<p>This protocol is from 'Flower Ca<sup>2+</sup> channel in CME and ADBE' of Yao CK et al.</p>
<div>
<div>
<p> </p>
<p>Please see the manuscript for the full method details.</p>
</div>
</div>
</div>

[BEFORE_START]
<p>You'll need: </p>
<p> </p>
<p><strong>0 mM Ca<sup>2+</sup> hemolymph-like (HL)-3 solution:</strong></p>
<ul>
<li>70 mM NaCl</li>
<li>5 mM KCl</li>
<li>10 mM MgCl<sub>2</sub></li>
<li>10 mM NaHCO<sub>3</sub></li>
<li>5 mM trehalose</li>
<li>5 mM HEPES (pH 7.2) </li>
<li>115 mM sucrose</li>
</ul>
<p> </p>
<p><strong>90 mM K<sup>+</sup>/0.5 mM Ca<sup>2+</sup> stimulation:</strong></p>
<p>(or alternative 60mM K<sup>+</sup>/1mM Ca<sup>2+</sup>)</p>
<ul>
<li>25 mM NaCl</li>
<li>90 mM KCl</li>
<li>10 mM MgCl<sub>2</sub></li>
<li>10 mM NaHCO<sub>3</sub></li>
<li>5 mM trehalose</li>
<li>5 mM HEPES (pH 7.2)</li>
<li>30 mM sucrose</li>
<li>0.5 mM CaCl<sub>2</sub></li>
</ul>
<p><strong>solution of 90 mM K<sup>+</sup>/2 mM Ca<sup>2+</sup>/200 mM chlorpromazine:</strong></p>
<ul>
<li>25 mM NaCl</li>
<li>90 mM KCl</li>
<li>10 mM MgCl<sub>2</sub></li>
<li>10 mM NaHCO<sub>3</sub></li>
<li>5 mM trehalose</li>
<li>5 mM HEPES (pH 7.2)</li>
<li>30 mM sucrose</li>
<li>2 mM CaCl<sub>2</sub> </li>
<li>200 mM chlorpromazine</li>
</ul>
<p> </p>

[STEPS]
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TITLE: No Title
AUTHORS: 

[DESCRIPTION]
For <a href="https://www.protocols.io/view/One-step-growth-curves-for-Cellulophaga-phages-ddh235" target="_blank">One-step growth curves for Cellulophaga phages</a> protocol and <a href="https://www.protocols.io/view/Transcriptomics-During-One-Step-Growth-Curves-for-dem3c5" target="_blank">Transcriptomics During One-Step Growth Curves for Cellulophaga Phages</a> protocol.

[STEPS]
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TITLE: Preparation of 0.1M Tris HCl, pH 6.4
AUTHORS: Nicola O'Reilly

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Purpose of examination / Clinical relevance</span></div><div class = "text-block">At the end of 2019, several pneumonia cases were reported in Wuhan, China and the pathogen was confirmed as a new viral strain. World Health organization has named the newly identified coronavirus as 2019-nCoV, also known as COVID19. The disease developed into a dangerous pandemic, posing major challenges to the NHS. Although more research is necessary to better understand the virus, in response to the emergency, simple and rapid testing is essential to identify the virus in infected individuals. This will aid the implementation of efficient interventions to contain the spread, and distinguish healthcare workers who have been infected, and are required to self-isolate, from those showing similar symptoms but which are not 2019-nCoV associated. The latter category may continue to work, alleviating stress on hard-pressed healthcare resources. 2019-nCoV is an RNA virus, and the diagnostic tests detect viral RNA in swabs from patient airways using a reverse transcriptase PCR assay. Samples are submitted to HSL, an accredited reporting laboratory, and transferred to the Crick for testing.</div><div class = "text-block"><span style = "font-weight:bold;">Principles of Examination</span></div><div class = "text-block"><span>This procedure involves the preparation of 0.1M Tris HCl pH 6.4 for making </span><a href="https://www.protocols.io/private/D1949D20846811EA9B0C0242AC110002" style = "text-decoration:underline;color:blue;cursor:pointer;"><span style = ":;">L6 virus inactivation buffer</span></a><span style = ":;">.</span></div></div>

[STEPS]
?. Please select between the recipes for 1 L 0.1M Tris HCl pH 6.4 and 5 L 0.1M Tris HCl pH 6.4.
Work in fume hood!
?. Weigh out  and add to a 1 L beaker.
[Tris HCl]
?. Measure out  and add to the beaker.
[milliQ water]
?. Add a magnetic flea and place on a magnetic stirring plate to mix the solution.
?. Add a newly calibrated pH meter into the solution to observe the pH.
?. Tune pH with .
[NaOH]
?. Transfer to a 1 L graduated cylinder and make up to 1000 ml using milliQ water. Swirl.
?. Vacuum filter into sterile bottles.
?. Label with batch number and date.
?. Store  at  ( – ).
[0.1 M Tris HCl, pH 6.4 solution]
0 Room temperature
15 °C
25 °C
?. Weigh out  and add to a 5 L beaker.
[Tris HCl]
?. Measure out  and add to the beaker.
[milliQ water]
?. Add a magnetic flea and place on a magnetic stirring plate to mix the solution.
?. Add a newly calibrated pH meter into the solution to observe the pH.
?. Tune pH with .
[NaOH]
?. Make up to 5 L using milliQ water by transferring to a second 5 L beaker via a 2 L graduated cylinder. Swirl.
?. Vacuum filter into sterile bottles.
?. Label with batch number and date.
?. Store ,  solution at  ( – ).
[Tris HCl]
0 Room temperature
15 °C
25 °C
TITLE: Nurses’, patients’, and informal caregivers’ attitudes toward aggression in psychiatric hospitals: a comparative survey study
AUTHORS: Maritta A Välimäki, Joyce       Y Lam, Daniel      Bressington, Teris  Cheung, Wai Kit Wong, Po Yee Ivy Cheng, Chi Fai Ng, Tony Ng, Chun Pong Yam, Glendy Ip, Lee Paul, Tella Lantta

[DESCRIPTION]

Attitudes toward aggression is a controversial phenomenon in psychiatry. This study examined and compared attitudes toward patient aggression in psychiatric hospitals from the perspectives of nurses, patients and informal caregivers and identified factors associated to these attitudes. A total of 2,424 participants completed a self-reported instrument regarding attitudes toward aggression (12-items Perception of Aggression Scale; POAS-S). We analyzed data from nurses (n = 782), patients (n = 886), and informal caregivers (n = 765). Pearson’s r correlations were used to examine associations between variables. Differences between group scores were analysed using ANOVA/MANOVA with post-hoc Sheffe tests. Multivariate logistic regression models and logistic regression analysis was used to examine the effects of respondents’ characteristics on their attitudes toward aggression. Nurses had significantly more negative and less tolerate perceptions toward aggression (mean [SD] 47.1 [7.5], p<0.001) comparing to patients (mean [SD] 44.4 [8.2]) and informal caregivers (mean [SD] 45.0 [6.9) regarding a POAS-S total score. The same trend was found in dysfunction and function sub-scores (mean [SD] 25.3 [4.1] and 15.0 [3.6], respectively; the differences between groups were in both cases both was statistically significant (p <0.001) comparing to patients (mean [SD] 23.7 [5.3] and 14.0 [4.1], respectively) and informal caregivers (mean [SD] 24.4 [4.2] and 13.9 [3.5], respectively). The study offers new understanding of aggressive behaviour in different treatment settings where attitudes toward patient behaviour raises ethical and practical dilemmas. These results indicate a need for more targeted on-the-job training for nursing staff, aggression management rehabilitation programs for patients, and peer-support programs for informal caregivers focused on patient aggression.

[STEPS]
1.
TITLE: Provasoli Enriched Seawater (PES) medium solution
AUTHORS: Anton Kuech

[DESCRIPTION]
This protocol outlines the steps required to prepare Provasoli Enriched Seawater (PES) medium in accordance with the method by Provasoli (1966).
 


[BEFORE_START]
It is important to follow the order of chemicals added as shown in the protocol.

[STEPS]
SECTION: Primary solution
3. 6000 mL   
SECTION: Primary solution
3.1. Add  23.4 g   (or 30 g   Tris buffer)
SECTION: Primary solution
3.2. Add 1500 mL  P II-solution   
SECTION: P II solution
1. 5000 mL    

SECTION: Primary solution
3.3. Add 1500 mL Fe-solution   
SECTION: Fe-solution
2. 5000 mL   

SECTION: Primary solution
3.4. Add 21 g   
SECTION: Primary solution
3.5. Add 2.12 g   

SECTION: Primary solution
3.6. Add 16 mg   
SECTION: Primary solution
3.7. Pasteurise for 120 min  at 99 °C 
SECTION: Vitamins
4. Vitamins to be added through a sterile single injection with a front filter (0.2 μm) in the cooled off primary solution (maximum temperature of 60 °C ).
SECTION: Vitamins
4.1. 30 mL  (10 mg per 500 ml  )
SECTION: Vitamins
4.2. 30 mL    (500 mg per 500 ml  )
SECTION: Vitamins
4.3. 30 mL   (5 mg per 500 ml   )
SECTION: Storage
5. Store PES in Duran bottles in a cold and dark setting at 10 °C .
 

SECTION: Application
6. Culturing medium is created by adding 200 mL PES to 10 L sterile seawater.
SECTION: P II solution
1.1. Add 5 g   
SECTION: P II solution
1.2. Add 5.7 g   

SECTION: P II solution
1.3. Add 0.62 g   

SECTION: P II solution
1.4. Add 0.11 g   

SECTION: P II solution
1.5. Add 0.024 g    

SECTION: P II solution
1.6. Add 0.245 g   

SECTION: Fe-solution
2.1. Add 3.3 g   

SECTION: Fe-solution
2.2. Add 3.51 g
TITLE: Determining biofilm growth amount (dry weight)
AUTHORS: Weizhuo.Chen, An.Huang

[DESCRIPTION]
This protocol will help determine the amount of biofilm grown on carriers.

[STEPS]
SECTION: IPTG induction
1. Escherichia coli grown overnight was diluted by LB to OD600=0.4-0.6.


SECTION: IPTG induction
2. IPTG was added to cell culture to 1mM IPTG finally, and incubated 3h at 171 rpm, 37℃ in orbital shaking incubator.
SECTION: Sample preparation
3. For each bottle:
(1) The total mass of thirty Moving Bed Biofilm Reactor (MBBR) carriers were measured in advance by balance. Record all weights.
(2) Cells culture, thirty MBBR carriers and silver nitrate (to 6μM) solution were added into each bottle.
(3) Put all bottles into a 37℃ incubator.
 

SECTION: Hot dry weight measurement
4. For each day,
(1) MBBR carriers were dried by oven at 60℃.
(2) Paddings were cooled to room temperature.
(3) Total mass of thirty MBBR paddings were measured.
Repeat steps above until weight did not change. Record the weight.
SECTION: Hot dry weight measurement
5. The variation of MBBR carrier mass could be calculated, thus indicating the accumulated biomass (biofilm) on the carriers.