eamag/protocols-with-steps huggingface dataset:
TITLE: VALAP protocol
AUTHORS: Elizabeth Fozo

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">VALAP is a biologically inert method for sealing coverslips, making it usefulfor live cell imaging.</div></div>

[STEPS]
?. [Use: Method 1 - "Painting" it on]
Put the jar or bottle on a hotplate on the lowest temperature setting.  Do not leave unattended!!!
?. [Use: Method 1 - "Painting" it on]
Wait for the Valap to melt fully.
?. [Use: Method 1 - "Painting" it on]
Using a cotton-tipped applicator, make sure it is well loaded with liquid Valap.
?. [Use: Method 1 - "Painting" it on]
Quickly wipe the Valap along the open edge of the coverslip, half on the coverslip and half on the slide.
?. [Use: Method 1 - "Painting" it on]
Repeat for the rest of the coverslip edges that need to be sealed.
?. [Use: Method 1 - "Painting" it on]
Be careful not to get Valap on microscope optics!  It is very difficult to remove and it's presence will significantly degrade image quality.
?. [Use: Method 2 - Careful placement]
This method uses a small metal spatula and an alcohol burner.
?. [Use: Method 2 - Careful placement]
Use the spatula to take a small amount of valap from the container.
?. [Use: Method 2 - Careful placement]
Hold it over the flame to melt it.
?. [Use: Method 2 - Careful placement]
Put a small bead of valap along the open edge of the coverslip.
?. [Use: Method 2 - Careful placement]
Repeat for the other sides of the coverslip
Both methods take a bit of practice to do quickly.  Method 2 creates a smaller profile and does not encroach on the coverslip surface as much as Method 1.
TITLE: Yale - Blood Urea Nitrogen
AUTHORS: John Stack, Gary Cline

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Summary: </span></div><div class = "text-block">Procedure used to measure the concentration of Blood Urea Nitrogen(BUN) in blood, plasma, and serum. Urea is determined by the enzymatically coupled reactions of urease (to form ammonia) and glutamate dehydrogenase (conversion of ammonia and glutamate to glutamine with oxidation of NADH to NAD). The rate of NAD formation is monitored by the change in absorbance at 340 nm. </div></div>

[STEPS]
?. Calibrate Cobas for BUN analysis by running a multi analyte standard and two assayed control serums.
?. Sample handling as performed by the Cobas Mira Plus.    a) Cobas pipettes 2 µL of sample into a cuvette slot.    b) Absorbance is measured at 340 nm.    c) Add 200 µL of BUN liquid reagent.    d) Mixture is incubated at 37ºC for 10 minutes.    e) Absorbance is measured at 340 nm. Change in absorbance is calculated.
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<div>
<div>
<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>
</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> </p>

[STEPS]
?. 
?. 
?. 
?. 
?. 
?.
TITLE: Library construction of metabarcoding at DNBSEQ-G400 with MGIEasy universal DNA library prep Kit
AUTHORS: Xiaohuan Sun, Yuehua  Hu, Zewei  Song

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Library construcion steps following MGIEasy universal DNA library prep set user manual started from end repair and A-tailing to single strand circularizaiton in chaptor 3. 500 ng of gDNA is used in this Library Construction Protocol. The steps were described bellow. </div><div class = "text-block">https://en.mgitech.cn/Uploads/Temp/picture/20191023/5db00efef2bd7.pdf</div></div>

[STEPS]
?. End-repair and A-tailingsteps are performed with ERAT Enzyme Mix in MGIEasy Universal DNA Library Prep Set (Cat. No: 1000006985).
?. Adaptor ligation step following the instruction bellow by using  MGIEasy DNA Adaptors.
5 mL
?. Cleanup of Adapter-ligated DNA by using DNA clean beads and freshly prepared 80% ethanol, final DNA was elute by  of TE buffer.
40 mL
?. of purified Adapter-ligated DNA sample with  PCR enzyme mix and  PCR primer mix were used for PCR amplification. We amplified samples using the following cycling conditions:  for ; 7 cycles of for ,  for , and  for ; and then a final extension at  for .
19 µl
25 mL
6 mL
98 °C
98 °C
60 °C
72 °C
72 °C
?. PCR products are cleaned by DNA clean beads.  of TE buffer is used to elute the DNA.
32 mL
?. Quantify the purified PCR products with dsDNA Fluorescence Assay Kits such as Qubit® dsDNA HS Assay Kit or Quant-iTTM PicoGreen® dsDNA Assay Kit. The required yield for PCR products is ≥ 1 pmol. For pooled sequencing, please follow instructions provided by MGIEasy DNA Adapters User Manual. The total yield after pooling should be 1 pmol, with a total volume ≤ .
48 µl
?. PCR products are stored at  for  for denaturation. After the reaction is complete, immediately place the PCR tube on ice for .
95 °C
?. Single strand circularization is prepared with single strand circularization reaction mixture consisting of  splint buffer and  DNA rapid ligase. The mixture is incubated at  for . The mixture should be immediately placed on ice after the completeness of the reaction.
11.6 µl
0.5 µl
37 °C
?. The circulized products are therefore used for sequencing at DNBSEQ-G400.
TITLE: Isolation of Nuclei from Frozen Human Peripheral Nerve
AUTHORS: Alexander Chamessian

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">This protocol details the efficient isolation of intact nuclei from frozen peripheral nerve from human (e.g. sciatic, tibial, etc.)</div></div>

[STEPS]
?. [Tissue Procurement]
Nerve samples should be collected as quickly as possible from the donor.Place the nerve in a clean, labeled container.Snap-freeze in liquid nitrogenStore at -80C until usage
?. [Cryo-Pulverization]
Remove the sample from -80C storage and maintain on dry ice. Do not let it thaw.
TITLE: Preparation of oxalate reagent
AUTHORS: Yingyu Hu, Zoe V Finkel

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">This protocol describes how to prepare oxalate reagent, which is used to remove surface adsorbed phosphorus, so that the intracellular phosphorus quotas in microalgae can be measured.</div></div>

[STEPS]
?. [10 M NaOH solution]
Add  MilliQ water in a 250 mL beaker.
50 mL
?. [10 M NaOH solution]
Weigh  NaOH and slowly pour into the beaker.
40 g
?. [10 M NaOH solution]
Use squeeze bottle to rinse the weighing boat and transfer rinse water into the same beaker.
?. [10 M NaOH solution]
Use glass rod to gently stir and fully dissolve NaOH.
The solution is very hot and corrosive. It can cause skin burns and eye damage.
?. [10 M NaOH solution]
Carefully transfer NaOH solution into 100 mL volumetric flask by using glass rod.
?. [10 M NaOH solution]
Rinse beaker with small amount of MilliQ water three times, transfer rinse water into the flask.
?. [10 M NaOH solution]
Mix the solution by gently shaking the capped volumetric flask and top to 100 mL with MilliQ water.
?. [10 M NaOH solution]
Transfer the prepared reagent into a 250 mL PP bottle.
?. [10 M NaOH solution]
Label the bottle with SDS pictogram.
?. [Oxalate reagent]
In a 1000 mL beaker with stir bar, add  MilliQ water.
600 mL
?. [Oxalate reagent]
Add  EDTA,  sodium citrate,   KCl and  NaCl into the beaker, stir until all ingredients are dissolved.
18.6 g
14.7 g
0.74 g
5 g
?. [Oxalate reagent]
NaOH is added dropwise to bring pH in between 6 to 7 by using a transfer pipet
?. [Oxalate reagent]
Add  oxalic acid to the solution, stir the mixture while heating.
12.6 g
?. [Oxalate reagent]
After oxalic acid is completely dissolved, stop heating and let it cool to room temperature. A water bath filled with tap water can be used to speed up cooling.
?. [Oxalate reagent]
Add  NaOH dropwise to bring pH to
?. [Oxalate reagent]
Top to 1 L in volumetric flask with MilliQ water.
?. [Oxalate reagent]
Filter oxalate reagent by rapid flow to a 1 L PP bottle.
?. [Oxalate reagent]
Label the bottle and keep it at .
0 Room temperature
TITLE: Western blotting for LRRK2 signalling in macrophages
AUTHORS: sherbst  

[DESCRIPTION]
This protocol describes the immunoblotting for components of the LRRK2 signalling pathway (LRRK2, LRRK2 pS935 and phospho-Rabs) using Invitrogen NuPage SDS-PAGE reagents and the BioRad Turbo Blot transfer system. 

[STEPS]
SECTION: Preparation of protein lysates
1. Culture cells as usual in a 12-well or 6-well plate. 
 

SECTION: Preparation of protein lysates
2. Wash cells once in PBS 
SECTION: SDS-PAGE
8. Prepare protein lysates by adding sample loading buffer and reducing agent (eg NuPAGE LDS sample buffer and reducing agent).
 

SECTION: Transfer
13. Transfer SDS gel into 20% EtOH while preparing the transfer stack. Make sure the gel is submerged. 
SECTION: Immunolabelling and detection
16. Transfer PVDF membrane into 5 % milk/TBS-T and block for 60 min at RT with constant shaking. 

 

SECTION: Preparation of protein lysates
3. Add 100 μl per 12-well or  200 μl per 6 well of lysis buffer containing protease and phosphatase inhibitors. Keep samples on ice from now on.

SECTION: Preparation of protein lysates
4. Scrape cells using a cell lifter and transfer lysate to a 1.5 ml Eppendorf tube
SECTION: Preparation of protein lysates
5. Vortex and incubate on ice for 10 min.

SECTION: Preparation of protein lysates
6. Clarify lysate by spinning at 17000 x g at 4 °C for 15 min

SECTION: Preparation of protein lysates
7. Transfer supernatant into a fresh 1.5 ml Eppendorf tube and store at -20 °C.
 

SECTION: SDS-PAGE
9. Denature samples at 80 °C for 8 min

SECTION: SDS-PAGE
10. Prepare the SDS gel (4-12% Bis-Tris Nu-PAGE gel) by assembling the apparatus, filling the tank with MES running buffer and flushing all the wells with MES running buffer. 
SECTION: SDS-PAGE
11. Spin down the samples quickly in case of condensation and load wells carefully. Include a broad range molecular weight marker (eg Broad molecular weight ladder, ab116028, Abcam). 
SECTION: SDS-PAGE
12. Run the gel at 180 V for 35 min. 

SECTION: Transfer
15. Transfer proteins using BioRad pre-programmed protocols: Mixed MW setting (2.5A for 7 min). 

SECTION: Transfer
14. Prepare the transfer stack according to the manufacturer's instructions.
SECTION: Immunolabelling and detection
17. Prepare primary antibodies in 5 % milk/TBS-T and incubate membranes in antibody solution at 4 °C    with constant shaking 


 


SECTION: Immunolabelling and detection
18. Wash membranes by shaking in TBS-T for 5-10 min 

SECTION: Immunolabelling and detection
19. Repeat the wash step 2x 
SECTION: Immunolabelling and detection
20. Repeat washing steps as above. 
SECTION: Immunolabelling and detection
21. Detect signal using appropriate detection equipment. 
SECTION: Optional: stripping and re-blot
22. Depending on the primary antibodies and the detection method used, it might be necessary to strip the blot using a Western Blot Stripping buffer (eg #T7135A, Takara Bio) for 10 min at RT and repeat the primary and secondary antibody incubation steps.
TITLE: Perfusion
AUTHORS: Molly Brennan, Olivier George

[STEPS]
?. WHOLE ANIMAL FIXATION VIA TRANSCARDIAL PERFUSIONList of Stuff Needed for Perfusion-4% Formaldehyde-0.9% Hep-saline-Bottles & pump-Surgical tool (scissors, clamps, bone rongeurs, spatula)-Vials-timer
?. Preparation-Switch waste flow to deposit into chemical waste bucket-Prepare perfusion pumps with 0.9% saline and 4% paraformaldehyde, check flow-Test system; check for flow and air bubbles
?. Anesthesia-Inject Clorohydrate 35 % into the left lower ventral abdomen-Wait approximately 5 min for anesthetic to take effect-Pinch the tip of the tail forcefully to test for reaction-Add additional amount of anesthesia if necessary
?. Perfusion Rates:Koob Lab—Pump runs in RPMS-Prefixative rinse -saline- 3 minutes @ max 90 RPM(i.e. ~24mls/min)-Formaldehyde- 5 minutes @ max 90 RPM(i.e. ~24mls/min)-Continue Form- 15 minutes @ 65 RPM(i.e. ~16mls/min)
?. Surgery and Fixation (do under hood or safe place)-Place animal on the downdraft, make a “V” shape above the genital-Lift skin upwards firmly using forceps, pinch a small hole into the abdomen, then cut along the “V” line-Continue to cut upwards to expose the diaphragm, and then cut through the diaphragm-Cut through the rib cage to expose the thoracic cavity-Clamp open excess tissue for better access of the heart-While holding the middle of the heart steady with forceps, make a cut slightly right to the apex of the heart (left ventricle), using one tip of a sharp scissors, then insert both tips of small scissors to make a bigger hole-Insert catheter needle through the cut, up to the aorta. Clamp the entrance
?. -Make a cut in the dark red-colored right atrium,-Circulate 0.9% saline, make sure fluid is flowing freely, until the limbs of the rat are discolored (3 min)-Turn off saline pump, and circulate 4% paraformaldehyde (approximately 250 ml) until neck is stiffened (3+15 min)
?. Brain Removal-Use a pair of scissors to cut connective tissue under the skin, reaching in between the eyes-Cut skin open, exposing muscles on top of skull to the eyes-Use scalpel, cut muscle connecting to the skull bone on both sides-Carefully place bone cutter against the inside of the skull, and make cuts following the scalpel incision-Lift the occipital, parietal, and frontal bones using forceps, fully exposing the brain-Cut membrane on top and scoop brain out of the skull; cut optical nerves and connective tissue-Store brain in 4% fixative for 1 day.
?. Sucrose Treatment-Take the brain out of the fixative, dab dry on utility towel-Place brain in 20% sucrose solution (in distilled water) until it sinks to the bottom of the vial-Place brain in 30% sucrose solution (in distilled water) for another day until it sinks to the bottom of the vial-Freeze brains before cutting in dry ice.
TITLE: URMC TriState SenNet Mouse Lung Digestion
AUTHORS: Gagandeep Kaur, Irfan Rahman

[DESCRIPTION]
The objective of this document is to share the material and kits used and steps involved in digesting the mouse lungs to perform scRNA analyses. This study was performed for the TriState SenNet TMC Bioanalyses Core at the University of Rochester, as part of the Cellular Senescence Network Program (SenNet).  This protocol ensures minimum damage to the cells in suspension and ensures maximum viability to conduct downstream analyses.

[STEPS]
SECTION: Lung Tissue Dissociation
2. 
Immediately transfer the tubes to MACS Tissue Dissociator and run user-defined program named m_lung_01_02 for mouse lungs.

SECTION: Lung Tissue Dissociation
1. Mince tissue finely, place into 50ml GentleMACS C-tube with liberase enzymatic cocktail .
SECTION: Lung Tissue Dissociation
3. Thereafter, place 50ml conical with digestion into an incubating rocker for 30 minutes at 37 °C . 
NOTE: If you still see chunks of lung tissue in the tubes after this step then transfer to the MACS Tissue Dissociator and run user-defined program named m_lung_02_01 for mouse lungs for 10-15 sec.
SECTION: Lung Tissue Dissociation
4. Remove conical from rocker and strain the cell suspension through 70 micron into 50mL tube 
SECTION: Lung Tissue Dissociation
5. Add 3 ml DMEM (10% FBS) through the strainer to collect the remaining cell suspension
SECTION: Lung Tissue Dissociation
6. After straining, centrifuge at 300g for 5 minutes at 4 °C.

SECTION: Lung Tissue Dissociation
7. Remove supernatant, add 500 µl of red blood cell lysis to cell pellet. Incubate the cell suspension on ice for 5 min.

SECTION: Lung Tissue Dissociation
8. After incubation add 4.5 mL of DMEM (10% FBS) to the suspension and centrifuge at 300g for 5 minutes at 4 °C .

SECTION: Lung Tissue Dissociation
9. Remove supernatant, re-suspend pellet in 2 mL DMEM (10%FBS)
SECTION: Lung Tissue Dissociation
10. Count cells and viability using AO/PI method, confirm reading under microscope
SECTION: Lung Tissue Dissociation
1.1. For 50mg tissue use 0.5 ml liberase + 2ml DMEM + 2ul 5 unit/ul DNase I.
TITLE: Protocols for Recinto et al. &#34;A rewiring of the earliest immune events leading to T-cell mediated disease following intestinal microbial infection  in a PINK1 KO mouse model of Parkinson’s disease&#34;
AUTHORS: Sherilyn Junelle Recinto, adam.macdonald, Moein Yaqubi, Alexandra Kazanova

[DESCRIPTION]
Our group has developed GI-targeted pathogen-induced PD mouse modeling systems (in PINK1 KO mice with gram negative bacterial infections) and found that T cells are a major player in driving PD-like motor symptoms at late stages following infection. Herein, we now map the initiating immune events at the site of infection at the earliest stages with the goal of shedding light on the earliest mechanisms triggering T cell-mediated pathological processes relevant to PD. Using unbiased single cell sequencing, we demonstrate that myeloid cells are the earliest dysregulated immune cell type in PINK1 KO infected mice (at 1-week post-infection) followed by a dysregulated T cell response shortly after (at 2 weeks post-infection). We find that these myeloid cells have an enhanced proinflammatory profile, are more mature, and develop enhanced capacity for antigen presentation. Using unbiased prediction analysis, our data suggests that cytotoxic T cells and myeloid cells are particularly poised for interacting with each other, and we identify possible direct cell-cell interaction pathways that might be implicated. 

[STEPS]
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p>The protocol describes guidelines and steps for taking repeated stabilometry measurements. The parameters calculated from repeated measurements can be user for reliability analysis of certain center of pressure parameters used to describe postural control and balancing capability. The guidelines are based on recommendations of the literature. It is important to be consistent during all the measurements and follow the guidelines as they regulate important aspects of the measurements. The changes of these aspects can signifficantly alter the motion of the center of pressure.</p>

[BEFORE_START]
<p>Register the basic anthropometrical data of the participants.</p>
<p>Determine the dominant leg by a balance recovery test.</p>
<p>The perturbation is a nudge from the tester applied to the subject at the midpoint between the scapulae from directly behind the subject and sufficient to require the participant to respond by taking a step. The leg that the subject uses to recover balance is considered to be the dominant leg. (Hoffman M, Schrader J, Applegate T, Koceja D. Unilateral postural control of the functionally dominant and nondominant extremities of healthy subjects. J Athl Train. 1998;33: 319–322)</p>

[GUIDELINES]
<p>Avoid audio or visual disturbance of the subjects during the trials, maintain comfortable temperature for the measurements.</p>
<p>Position the investigated subjects in bipedal standing, with the distance between the 2 ankle joint centers equal to the distance between the right and left anterior superior iliac spines.</p>
<p>Maintain both limbs in full knee extension, the heels aligned in a line, and the feet parallel and faced forward, with arms resting by the sides.</p>
<p>During the stance on the dominant leg, the dominant leg should be in full knee extension, and the non-dominant leg flexed at 90 degrees.</p>
<p>Since a 60 second single leg standing trial is challenging, in case the subject have to put down the other leg or shows substantial arm or whole body movements, the measurement should be aborted and repeated.</p>
<p>The exact foot placement should be maintained during the trials and visually controlled since its changes could affect the stabilometry parameters.</p>
<p>During stances with the eyes open, the eyes should be focused on the wall at eye level at least 3 meter from the subject.</p>
<p>The measurements can be started after the initial transients of the subject’s self-adjustment. The adjustment time is usually approximately 10 seconds, but a minimum of 5 second wait is required after the subject stood on the platform.</p>
<p>For the reliability assessment the measurements should be performed by the same person.</p>
<p> </p>
<p>Record 60 second trials (started after the initial transitions) on 100 Hz sampling frequency.</p>
<p>Apply a Butterworth low-pass digital filter on the center of pressure coordinates with 10 Hz cut-off frequency before calculating the center of pressure parameters.</p>
<p> </p>
<p>Generally, maintain the exact conditions under each trial of the subjects.</p>

[STEPS]
?. 
?. 
?. 
?. 
?.
TITLE: Dot Blots Analysis in the Separation of Anti-HIV Antibodies.
AUTHORS: Angel  Justiz-Vaillant

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Dot blot frequently is underutilized or is not frequently used because it cannot assess the molecular weight of macromolecules, but it is simple to perform and is useful when a particular biomolecule needs to be identified. In this protocol dot blot analysis is used for detection of anti-HIV antibody [1], and it is also available for determination or separation of any protein. It can be very useful in the immunodiagnosis of infectious diseases caused by viruses [1] and bacteria [2] and in addition for the determination of specific DNA sequences by using probes [3]. </div><div class = "text-block">References</div><div class = "text-block">1. Justiz Vaillant AA, McFarlane-Anderson N, Akpaka PE, Smikle MP, Ramirez N, et al. (2013) Use of Dot Blots Analysis in the Separation of Anti-HIV Antibodies in Animals. J Chromat Separation Techniq 4: 181. doi:10.4172/2157- 7064.1000181</div><div class = "text-block">2. Cardona-Castro N, Gotuzzo E, Rodriguez M, Guerra H. Clinical application of a dot blot test for diagnosis of enteric fever due to Salmonella enterica serovar typhi in patients with typhoid fever from Colombia and Peru.Clin Diagn Lab Immunol. 2000;7(2):312-313. doi:10.1128/cdli.7.2.312-313.2000</div><div class = "text-block"><span>3. Clément G, Benhattar J. A methylation sensitive dot blot assay (MS-DBA) for the quantitative analysis of DNA methylation in clinical samples.</span><span style = "font-style:italic;">J Clin Pathol</span><span>. 2005;58(2):155-158. doi:10.1136/jcp.2004.021147</span></div></div>

[STEPS]
?. Two (2) µl of 1:2 dilutions of chicken IgY or 2 µl of serum from rats and cats is dotted onto nitrocellulose paper. This animal specimens tested positive for the presence of anti-HIV antibodies by ELISA, after being immunized with HIV proteins.
?. Place the nitrocellulose membrane in a BioDot SF apparatus (Bio-Rad Laboratories, Richmond, CA, USA).
?. The membrane is blocked with 5 μL/well of fetal bovine serum with 1%Tris buffer saline.
?. Then 5 μL of a commercial conjugate (peroxidase-labeled HIV proteins; Murex Diagnostics, Norcross, USA) is added and allow to drain by gravity.
?. Finally, 5 μL of the substrate 3,3’,5,5’-tetramethylbenzidine (Sigma Chemical Co., St Louis, MO, USA) is added and the mixture is incubated for 20 min.
?. The reaction is stopped by washing the wells with distilled water under a vacuum.
?. The membrane is left to dry.
?. Visualization of color development.
TITLE: POLE TEST PROTOCOL
AUTHORS: Michael Lee

[DESCRIPTION]
This protocol details pole test.

[GUIDELINES]
Detailed records should be kept on every experiment, noting date, timing, subject test order, procedural details followed and any relevant observations pertaining to the assay (e.g. specific behavior or procedural errors).

[STEPS]
SECTION: Test
1. Place animal on horizontal pole facing the top.
SECTION: Test
2. Place pole upright with mouse facing upward.
SECTION: Test
3. Record the time to turn (orient facing down), and time to reach the bottom of the pole.
SECTION: Test
4. Maximum duration 2 min.

SECTION: Test
5. All animals tested for 3 trials with a minimum of 20 min between trials.
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p>Uses a custom Centrifuge pipeline to assign taxonomy to gene calls. </p>

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