eamag/protocols-with-steps huggingface dataset:
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<br />[Reference: GeneMATRIX Basi DNA Purification Kit, version 1.0, EURx company]</p>

[STEPS]
?. 
?. 
?. 
?. 
?.
TITLE: Pharmacological management and potentially inappropriate prescriptions for patients with acne
AUTHORS: Jorge  Machado Alba

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><span style = "font-style:italic;">Introduction</span><span>. Acne is a chronic inflammatory disease that involves the pilosebaceous follicle and mainly affects adolescents and young adults. Its pharmacological treatment involves topical and systemic medications, but a heterogeneous group of drugs may exacerbate or induce skin lesions. The aim was </span><a href="" style = "text-decoration:underline;color:blue;cursor:pointer;"><span style = ":;">to identify the pharmacological management and medications related to the exacerbation of skin lesions in patients diagnosed with acne</span></a><span>. </span></div><div class = "text-block"><span style = "font-style:italic;">Methods</span><span>. This was a cross-sectional study that identified the outpatient medication prescription patterns of patients with acne from a dispensing database of 8.5 million members of the Colombian Health System. Sociodemographic and pharmacological variables and the identification of prescriptions that were potentially inappropriate due to the risk of worsening acne were considered. </span></div><div class = "text-block"><span style = "font-style:italic;">Results</span><span>. A total of 21,604 patients with acne were identified. Their median age was 20.8 years [interquartile range: 17.3-27.3 years], and 60.7% were women. Treatment mainly involved antibiotics (79.9% of patients), especially doxycycline (66.0%) and retinoids (55.7%). A total of 17.2% of the patients had potentially inappropriate prescriptions, predominantly the use of progestogens with androgenic properties (8.9%). Women (OR:3.55; 95%CI:3.24-3.90) and patients with pathologies such as systemic lupus erythematosus (OR:18.61; 95%CI:7.23-47.93) and rheumatoid arthritis (OR:10.80; 95%CI:5.02-23.23) were more likely to receive inappropriate prescriptions, and the risk increased with each year of life gained (OR:1.02; 95%CI:1.02-1.03).</span></div><div class = "text-block"><span style = "font-style:italic;">Conclusion</span><span>. Patients with acne are excessively treated with systemic antibiotics, counter to clinical practice guidelines. Approximately one-fifth of these patients received some potentially inappropriate medication that could exacerbate their skin lesions.</span></div></div>

[STEPS]
TITLE: Preparation of Competent Cells of M. bovis BCG
AUTHORS: Jonathan Rey A Ibale

[DESCRIPTION]
This protocol is a standardized protocol obtained from the Polytechnic University of Hong Kong and is in current use in SC 193 Laboratory under Prof. Au Wing Ngor for M. bovis BCG experimentation. The cells produced thereof are electrocompetent. 

[STEPS]
2. Inoculate a starter bacterial stock to 100 mL 7H9 broth (0.1g casitone, 0.44 g sodium pyruvate) incubate in 37 °C  for 7-8 days without shaking. (Prolonged incubation causes dehydration so seal the flask/tube during incubation.)
3. Aliqout 1 mL from the primary culture into a fresh, pre-warmed 100 mL 7H9 broth and incubate in 37 °C  for 8 days, followed by another 8 days of incubation with shaking at 90 rpm. Measure absorbance in 600nm (OD600) before shaking and periodically during dynamic incubation phase.

1. Prepare a fresh 7H9 broth for every culture step. The 7H9 broth is prepared as follows:
1. Mix together:
0.47 g 7H9 broth powder
0.1 g casitone (pancreatic digest of casein)
0.40 g sodium pyruvate
200 µL glycerol
90 mL nuclease-free double-distille water (MQ water)
2. Autoclave mixture at 121 °C for 10 mins, and shake media gently after.
3. Cool to about 45 °C , then add 10 mL OADC and 1 mL 10% sterile Tween-80. (Sterile 10% Tween-80 is prepared by diluting 1 mL Tween-80 in 9mL sterile MQ water, then sterilized by filtration in 0.22µm membrane.)
4. The 7H9 broth may be stored in 4 °C  after preparation and can be used for up to two weeks.

4. When the absorbance OD600 reaches 0.4-0.5, add 0.1 volume of autoclaved 2M glycine, and incubate for another 16-24 hours. Perform a contamination check along with this incubation using BHI media pre-warmed and incubated in 37 °C . If the inoculation develops turbidity overnight, the culture is contaminated with non-specific bacteria or rapidly growing mycobacterium (RGM).
5. 
Aliquot bacterial suspension in 4 50-mL conical tubes (around 25 mL each) and centrifuge at 3000xg for 10 mins in RT. Remove media and wash cells, i.e. resuspend, centrifuge, then remove media, three (3) times with autoclaved and pre-warmed 10% glycerol first at 20 mL, 10 mL, then 5 mL. Resuspend the final pellet in 500 µL 10% glycerol (2% aliquot volume) and aliquot 200µL into sterile propylene tubes for storage in -80 °C .
TITLE: Impact of CBR
AUTHORS: Dr.  Md. Zahid Hossain, Dr. Kazi Md. Amran Hossain, Dr. Md. Feroz  Kabir

[DESCRIPTION]
Background
Community-Based Rehabilitation (CBR) is reaching the rehabilitation services to the doorstep of the stakeholders. The stakeholders include the persons with disabilities, their family members or caregivers including the society where they live. 10-15% of the population in Bangladesh has disabilities and 80% of them reside in rural areas. There is no exact prediction of the necessity of community-based rehabilitation in Bangladesh. To our best knowledge, there is a scarcity of data in this field. Moreover, the study recommends the implementation of CBR in the primary healthcare services in Bangladesh and evaluates them in the country's context. Our study is expected to meet the inquiries in a small-scale population of a selected district in Bangladesh. 

Objectives  
The main objective of the study will be to evaluate the outcome of Community Based Rehabilitation (CBR) in the Jashore District in Bangladesh. The specific objectives are to screen the community dwellers for the necessity of CBR, to detect the risk of disability (induced by Low back Pain), to elicit the impact of CBR on Risk reduction and pain remission on chronic painful Musculoskeletal impairments, and to find out the effectiveness of CBR on the health matrix in chronic neurological disabilities in children and adults. 

[BEFORE_START]
NA

[GUIDELINES]
The intervention protocol is provided in detail in the attached document 

[STEPS]
SECTION: Screening Camps
1. Screening Camps 
SECTION: Screening Camps
2. Inclusion according to eligibility
SECTION: Pretest Measures
3. Pretest Measures through Baseline measures 
SECTION: Pretest Measures
4. Primary Intervention 
SECTION: Pretest Measures
5. Immediate Posttest (if required)
SECTION: Intervention
6. Customized intervention for 12 sessions in 4 weeks 
SECTION: Posttest measures
7. Posttest measures through outcome tools 
SECTION: Follow Up
8. Follow up measures
TITLE: Dye-terminator DNA sequencing
AUTHORS: Diep Ganguly

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">This protocol (based on the BigDye® Terminator v3.1 Cycle Sequencing Kit) is for performing terminator cycling sequencing reactions for Sanger sequencing of amplified PCR products or plasmid DNA on the 3130X genetic analyser (Applied Biosystems).</div></div>

[STEPS]
?. [Enzymatic PCR clean-up]
If sequencing a PCR amplified DNA fragment, gel purify target DNA band based on expected fragment size (if multiple bands present). Perform gel purification with Wizard SV Gel and PCR Clean-Up System (Promega, as per attached Manufacterer's instructions) followed by enzymatic clean-up (hydrolyze excess primers and nucleotides) with the following reaction:  AB1ComponentVolume (µl)210X Antarctic phosphatase reaction buffer13Antarctic phosphatase0.54XRN-10.55Purified DNA fragment50-150 ng DNA6Nuclease-free waterto 10 µlIncubate the above in a thermal cycle for:37 °C for 30 minutes80 °C for 15 minutes.
AB1ComponentVolume (µl)210X Antarctic phosphatase reaction buffer13Antarctic phosphatase0.54XRN-10.55Purified DNA fragment50-150 ng DNA6Nuclease-free waterto 10 µl
?. [Terminator cycling reaction]
Perform sequencing PCR in PCR tubes (or 96-well plate) with BigDye Terminator cycling kit and forward or reverse primers.  AB1ComponentVolume (µl)2v3.1 Ready reaction mix135X Sequencing buffer1.5420 µM F/R Primer0.55Template (plasmid or cleaned PCR product)50-150 ng DNA6Nuclease-free waterto 10 µl5x reaction buffer=400 mM TRIS, 10 mM MgCl2
AB1ComponentVolume (µl)2v3.1 Ready reaction mix135X Sequencing buffer1.5420 µM F/R Primer0.55Template (plasmid or cleaned PCR product)50-150 ng DNA6Nuclease-free waterto 10 µl
?. [Terminator cycling reaction]
Run the following thermal cycling protocol:1 min at 96 °C30-40 cycles: 96 °C for 10 seconds, 50 °C for 5 seconds, and 60 °C for 4 min.Hold at 4-12 °C.
?. [Purification]
Transfer PCR reaction to nuclease-free eppendorf tube. To the reaction, add 2.5 µL of 125 mM EDTA (make sure it touches bottom of tube).
?. [Purification]
Add 30 µl of 100% ethanol, mix well (inversion).
?. [Purification]
Incubate at room temperature for 15 minutes.
?. [Purification]
Centrifuge at 4 °C at max speed for 30 minutes.
?. [Purification]
Discard supernatant and add 50 µl of ice-cold 70% ethanol.
?. [Purification]
Centrifuge at 4 °C at max speed for 5 minutes.
?. [Purification]
Discard supernatant and allow to air-dry in the dark for >15 minutes.
?. [Prepare for sequencing]
Resuspend the pellet (likely transparent) in 7.5 µL HiDi Formamide (add to any empty wells). Incubate at RT for 5 minutes then transfer to plate. Spin down briefly.
?. [Prepare for sequencing]
Incubate plate at 95 °C for 3 minutes (denature) then place immediately on ice. Spin down briefly.
?. [Prepare for sequencing]
Submit for sequencing on 3130X genetic analyser (Applied Biosystems). Keep samples on ice.
TITLE: Basic Protocol 3: Testing auxin-mediated degradation of the AID-tagged protein
AUTHORS: Kizhakke Mattada Sathyan, Thomas G. Scott, Michael J. Guertin

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Please refer to the description section of the protocol collection.</div></div>

[STEPS]
?. Make  auxin in water, aliquot and store at . The auxin is stable at  for several months. Use a fresh aliquot each time and do not refreeze.
-20 °C
-20 °C
?. Seed 6-well plates with AID-tagged cells to achieve ~75% confluent cells the following day (plate approximately 7-8x105 HEK293T cells).
?. Add a final concentration of  auxin dropwise to the media all over the plate and mix by moving the plate forward and backward and sideways. Do not swirl the plate.
?. Collect cells at regular intervals starting from no auxin treatment. Initially, we collect at 15 minutes, 30 minutes, 1, 2, 3, and 4 hours. Remove  media from each well and collect cells by pipetting up and down in the remaining media. Put the cells .
1 ml
on ice
?. Centrifuge cells immediately after collection using a fixed angle rotor table top centrifuge at  for  at  and carefully remove media by using a pipette.
Centrifuge: 6000 34
4 °C
Keeping cells on ice and centrifugation at 4°C significantly reduce further degradation of the tagged protein.
?. Add  2x SDS sample buffer directly into the pellet and pipette up and down several times. The lysate becomes highly viscous.
200 µl
Alternatively, directly add the 2x SDS sample buffer into the plate after washing with PBS and lyse the cells. Collect the lysate with a pipette.
?. Heat denature protein at  for , vortex for  and denature again for another . Store the lysate at .
95 °C
-20 °C
?. Check the auxin-induced degradation of the tagged protein (Figure 3C-E, 4B&C). Serially dilute the untreated lysate to ensure that the query bands of the Western are within the linear range of the assay. Load the serial dilution of the untreated AID-tagged control lysate and include the treated cell lysate. Continue with Western blotting using antibodies directed against the AID-tagged protein. The degradation of the AID-tagged proteins starts immediately after adding auxin. Determine the rate of degradation by plotting the measured intensity of the AIDtagged protein bands using densitometry and fit the data using nonlinear regression and a one-phase decay equation.
Although the AID system works in many cell types and organisms, each cell type and organism is unique and the co-factors of the ubiquitin system may be differentially active.
TITLE: BAF_Protocol_006 On-Bead Peptide Cleanup (Digested by Other Method) 
AUTHORS: nesf

[DESCRIPTION]
This protocol is for using beads as a cleanup step after protein is digested using a different method (not directly on  Sera-Mag beads), for example solution digest. The peptides are precipitated onto the beads to allow for removal of salts and detergents that will interfere with downstream mass spectrometry. 

[STEPS]
SECTION: On-Bead Clean-up
1. Stock Solution of magnetic Sera-Mag beads is made in section 1 of BAF_Protocol_004 and can be stored at 4C indefinitely and used as needed (never freeze).
SECTION: On-Bead Clean-up
2. Peptide digest (for example from a solution or in-gel digest) should be reduced in volume so that a 95% ACN (acetonitrile) final concentration can be made within the Eppendorf tube volume. This will precipitate the peptides onto the beads for clean-up. See next steps.
SECTION: On-Bead Clean-up
3. To the peptide digest that has been reduced in volume, add 2uL of bead stock mix (20ug).
SECTION: On-Bead Clean-up
4. Add 100% ACN to a final concentration of 95% ACN. Incubate for 8 minutes at room temperature.
SECTION: On-Bead Clean-up
5. Placed tubes on a magnetic rack for 2 minutes. Remove and discard supernatant.
SECTION: On-Bead Clean-up
6. Rinse beads with 180uL of 100% ACN, incubate for 30s then remove and discard supernatant.
SECTION: On-Bead Clean-up
7. Add 10uL of 2% DMSO in water. Give quick spin (2-3s) in bench-top centrifuge to aid liquid removal from tube walls to bottom of tube.
SECTION: On-Bead Clean-up
8. Place tubes in magnetic rack for 2 minutes and recover supernatant making sure not to recover any beads.
SECTION: On-Bead Clean-up
9. Dilute samples with 0.1% FA (formic acid in water) to <1% DMSO. Further clean-up using C18 tips may be done using BAF_Protocol_003.
TITLE: Background control of Gotcha RCA
AUTHORS: Chia-Hsien  Shih

[DESCRIPTION]
This protocol aims to verify that GotCha design works as expected.

[STEPS]
SECTION: Preparation
1. Add 5 µL of functional beads(Gotcha) into eppendorf

SECTION: Preparation
2. Centrifuge for 15000 rpm, 5 min and remove supernatant. Make sure that eppendorf should put on DynaMag when removing supernatant.

SECTION: Protocol of Control group without miRNA
3. Add 3 µL of 10X phi29 polymerase reaction buffer into eppendorf with functional beads

SECTION: Protocol of Control group without miRNA
4. Add 23.4 µL RNase-free water

SECTION: Protocol of Control group without miRNA
5. Add 3 µL of 2mM dNTPs

SECTION: Protocol of Control group without miRNA
6. Add 0.6 µL of 10U/ul phi29 polymerase

SECTION: Protocol of Control group without miRNA
7. Pipetting to mix well
SECTION: Protocol of Control group without miRNA
8. Incubate for 120 min at 31 Room temperature

SECTION: Protocol of Control group without miRNA
9. Add 1.5 µL of 20X evagreen dye 

SECTION: Protocol of Control group without miRNA
10. Add 1.5 µL of 0.5M EDTA buffer to suspend the reaction

SECTION: Measuring
18. Load 20 µL of reaction solution into 384-well plate

SECTION: Measuring
19. Measure the fluorescence excitation and emission intensity
SECTION: Protocol of Control group without phi29 polymerase
11. Add 3 µL of 10X phi29 polymerase reaction buffer into eppendorf with functional beads

SECTION: Protocol of Control group without phi29 polymerase
12. Add 21 µL RNase-free water
SECTION: Protocol of Control group without phi29 polymerase
13. Add 3 µL of 2mM dNTPs
SECTION: Protocol of Control group without phi29 polymerase
15. Pipetting to mix well
SECTION: Protocol of Control group without phi29 polymerase
14. Add 3 µL of 100nM miRNA

SECTION: Protocol of Control group without phi29 polymerase
16. Add 1.5 µL of 20X evagreen dye 

SECTION: Protocol of Control group without phi29 polymerase
17. Add 1.5 µL of 0.5M EDTA buffer to suspend the reaction
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p>Evaluación de la capacidad de neutralización del efecto del Lipopolisacarido (LPS) de inducir la producción de Factor de Necrosis Tumoral (TNFalfa) en Células Mononucleares de Sangre Periférica (PBMCs) tratadas con peptidos cationicos.  </p>
<p> </p>
<p>La realizacion de este protocolo fue posible gracias al apoyo del departamento administrativo de ciencia tecnología e innovacion, Colciencias a traves del proyecto 111356933173 convocatoria569-2012.</p>

[BEFORE_START]
<p>Preparar todos los reactivos y soluciones de trabajo con un día de anticipación.</p>
<p>Se recomienda realizar el ELISA de los sobrenadantes inmediatamente despues de colectados; para esto se debe tener en cuenta que la placa debe sensibilizarse un día antes del desarrollo del ELISA. </p>
<p> </p>

[GUIDELINES]
<p>La capacidad de los peptidos para neutralizar o inhibir la producción de Lipopolisacarido (LPS) es medida a traves de la capacidad de los mismos de inhibir la producción de TNF alfa en Células Mononucleares de Sangre Periferica (PBMCs). El cultivo de PBMCs fue realizado con 250.000 células por pozo en un plato de cultivo de 96 pozos  (Cellstar, greiner bio-one, cat-N°655180), las celulas fueron tratadas por triplicado con 10 ng/mL de LPS  (<em>Escherichia coli, </em>LPS 026:B6, Sigma Aldrich L2654), se utilizan controles por triplicado con medio de cultivo y los tratamientos con diferentes concentraciones del péptido a evaluar con LPS y sin LPS. </p>
<p> </p>
<p> </p>
<p>Se incuban las células por 12 horas (este tiempo se debe estandarizar) a 37 ºC y 5 %CO<sub>2; </sub>el sobrenadante es colectado y reemplazado con medio de cultivo celular con 44 µM de Rezasurin con el fin de evaluar la viabilidad celular. Posterior a esto los sobrenadantes colectados se les mide la producción de TNFalfa por ELISA (Human TNF-ɑ ELISA MAX, BioLegend), siguiendo las instrucciones del fabricantes. </p>
<p> </p>

[STEPS]
?. 
?. 
?. 
?. 
?.
TITLE: nCoV-2019 McGill RT Protocol, Lunascript
AUTHORS: Shu-Huang Chen, Sarah Reiling, Josh Quick

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Artic nCoV-2019 McGill modified Lunascript Reverse Transcriptase sequencing protocol.</div></div>

[STEPS]
?. [cDNA preparation]
Mix the following components in a 0.2 mL 8-strip tube; Component                                        VolumeLunaScript RT SuperMix (5x)          Nuclease-free water                         Template RNA                                   Total
4 µl
5 µl
11 µl
20 µl
Viral RNA input from a clinical sample should be between Ct 18-35. If Ct is between 12-15, then dilute the sample 100-fold in water, if between 15-18 then dilute 10-fold in water. This will reduce the likelihood of PCR-inhibition.
A mastermix should be made up in the mastermix cabinet. Tubes should be wiped down when entering and leaving the mastermix cabinet.
?. [cDNA preparation]
Gently mix by pipetting and pulse spin the tube to collect liquid at the bottom of the tube.
?. [cDNA preparation]
Incubate the reaction as follows: for  for  for Place on ice for
25 °C
55 °C
95 °C
[ or store cDNA at -20C]
TITLE: Preparation of dsRNA viruses for next-generation sequencing
AUTHORS: Alexander H Wilcox, Erik Delwart, Samuel L Díaz Muñoz

[DESCRIPTION]
<div class = "text-blocks"></div>

[STEPS]
?. Viral lysates should be passed through a 0.22μm filter to remove host debris, then treated with nucleases to degrade extracapsular nucleic acids.  We added 25μl DNAse I, 50μl RNAse A/T1 and 1X DNAse I Buffer to 1ml filtrate, and incubated for 1 hour 30 minutes at 37ºC.
?. RNA should be extracted using a commercially available kit.  We used an RNeasy Mini Kit (Qiagen).  Kits that require carrier RNA are not recommended because this will interfere with sequencing.
?. Quantify the RNA using Qubit.  Note that low viral titres and/or small viral genomes may result in the amount of RNA being below the Qubit's limit of detection.
?. Add 50% DMSO to RNA sample, and incubate for 1 hour 30 minutes at 65ºC.  Purify RNA using a column cleanup kit (we recommend Qiagen's RNeasy MinElute Cleanup Kit) or your preferred other method (e.g. ethanol precipitation, TRIzol purification).
?. First strand synthesis.  Add 5μl purified RNA to 1μl random hexamers, 1μl dNTPs and 3μl nuclease-free water, and incubate for 5 minutes at 65ºC.  Place on ice for 1 minute, then add 0.01M DTT, 5mM MgCl2, 1X reverse transcriptase buffer, 1μl RNAseOUT and 1μl Superscript III enzyme to each reaction, using nuclease-free water to bring reactions to a total volume of 20μl.  Incubate in a thermal cycler for 10 minutes at 25ºC, 50 minutes at 50ºC and 5 minutes at 85ºC.  Place reactions on ice.
?. Second strand synthesis.  Add 1μl dNTPs, 0.5μl DNA ligase, 2μl DNA polymerase I, 0.5μl RNAse H, 1X second strand synthesis buffer, and use nuclease-free water to bring the total volume to 40μl.  Incubate for 5 hours at 16ºC, and purify DNA using your preferred method (we used a Macharey-Nagal Nucleospin Gel and PCR Clean-up kit).  Quantify DNA using Qubit.
?. Prepare DNA libraries for sequencing using your chosen platform and library preparation kit (we used Nextera XT).
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<div class="page" title="Page 12">
<div class="layoutArea">
<div class="column">
<div class="column">The steps for preparing the lysate are different depending on the starting material. Please ensure you follow the correct procedure for your starting material (see the section <a href="https://www.protocols.io/view/genomic-dna-removal-and-total-rna-purification-fro-f48bqzw?guidelines" target="_blank">Disrupting and Homogenizing Starting Materials</a> and the different lysate preparation protocols <a href="https://www.protocols.io/view/isolate-ii-biofluids-rna-kit-f5kbq4w" target="_blank">here</a>). The subsequent steps detailed in <a href="https://www.protocols.io/view/genomic-dna-removal-and-total-rna-purification-fro-f48bqzw" target="_blank">Genomic DNA removal and total RNA purification</a> are the same in all cases.</div>
<div class="column"> </div>
<ul>
<li>
<p>This protocol is designed for the isolation of all sizes of RNA (including miRNA) from a wide variety of biofluids derived from all human and animal sources collected with nasal or throat swabs.</p>
</li>
<li>
<p>Follow this protocol for isolating RNA from viruses collected with nasal or throat swabs.</p>
</li>
</ul>
</div>
</div>
</div>

[BEFORE_START]
<div>
<div>
<div>
<div class="b-start-block editable-content">
<div class="page" title="Page 12">
<div class="layoutArea">
<div class="column">
<ul>
<li>Ensure Lysis Buffer RX is prepared (see guidelines).</li>
<li>Ensure that all solutions are at room temperature before use.</li>
<li>Two types of spin columns are provided with this kit: the <strong>Genomic DNA Removal Colum </strong>(blue ring) and the<strong> RNA Column</strong> (black ring). Ensure the correct column is used for each step of the procedure.</li>
<li>All centrifugation steps are carried out in a benchtop microcentrifuge at 14,000 x g except where noted. Perform all centrifugation steps at room temperature.</li>
<li>It is important to work quickly during this procedure.</li>
</ul>
</div>
</div>
</div>
</div>
</div>
</div>
</div>

[GUIDELINES]
<p>Please review the Guidelines under <a href="https://www.protocols.io/view/genomic-dna-removal-and-total-rna-purification-fro-f48bqzw?guidelines" target="_blank">Genomic DNA removal and total RNA purification from all types of lysate</a> for important details.</p>

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