eamag/protocols-with-steps huggingface dataset:
TITLE: Immunohistochemistry free-floating rat brain cryosections  
AUTHORS: mariangela.massarocenere

[DESCRIPTION]
Protocol for  immunohistochemistry on rat brain cryosections 

[STEPS]
SECTION: 1. Sections selection
1. Collect the cryosections needed for a caudo-rostral representation of each brain region (every fifth  or sith sections depending on the section's thickness, brain region, and animal species) into a 24-well-plate (3-4 sections per well)
SECTION: 2. Inactivation of endogenous peroxidase
2. Incubate sections with endogenous peroxidase blocking solution: PB1X + 3% H2O2 (500 µ l/well) for at RT

SECTION: 1. Sections selection
1.1. Wash in PB1X 3 times x 5 min  (500 ul) at RT
SECTION: 2. Inactivation of endogenous peroxidase
2.1. Wash in PB1X 2 times x 5 min  (500 ul) at RT
SECTION: 3. Blocking
3. Incubate with blocking solution (500 µ l/well) : PB1X + 10%NGS (serum from the same species as the host of the secondary antibody) + 0.3% TritonX-100, at RT
SECTION: 4. Primary antibody incubation
4. Remove the blocking solution and incubate sections with PB1X+ 5%NGS+0.3% TritonX-100+ primary Ab (500 µ l/well) for 24/72 h, depending on the Ab, at +4°C 
SECTION: 4. Primary antibody incubation
4.1. Wash  in PB1X 3 times x 5 min  (500 ul) at RT 
SECTION: 5. Secondary biotinylated antibody incubation
5. Incubate sections with adequate secondary biotinylated Ab diluted in PB1X+ 0.3% TritonX-100, at RT 
SECTION: 5. Secondary biotinylated antibody incubation
5.1. Wash n PB1X 3 times x 5 min  (500 ul) at RT 
SECTION: 6. Extravidin-peroxidase reaction
6. Incubate with Extravidin-peroxidase buffer solution diluited in PB1X+0.3% TritonX-100, at RT
SECTION: 6. Extravidin-peroxidase reaction
6.1. Wash n PB1X 3 times x 5 min  (500 ul) at RT
SECTION: 7. Substrate preparation
7. Allow each DAB tablet to reach room temperature before use and  Dissolve a DAB tablet in 15 mL of Tris-buffered saline, pH 7.6
SECTION: 7. Substrate preparation
7.1. Add 12 µL of fresh 30% hydrogen peroxide prior to use
SECTION: 7. Substrate preparation
7.2. Filter the solution through a 0.2 µm filter immediately before use 
SECTION: 8. Development
8. Add 500 µl/ well of substrate solution in the dark 
SECTION: 8. Development
8.1. Remove and clean al material with bleach 
SECTION: 8. Development
8.2. Wash 2 times x 5 min in PB1x 
SECTION: 9. Section Mounting
9. Mount sections on SuperFrost plus slides and let it dry overnight 
SECTION: 10. Dehydratation
10. Dehydrate in consecutive steps of ethanol solutions (50%, 70%, 95%, 100%)
SECTION: 10. Dehydratation
10.1. Incubate slides in 2 times x 5 min in Xylene
SECTION: 11. Mount Coverslips
11. Coverslip slides with mounting medium, remove bubbles if any, and let dry
TITLE: DNA barcoding SOPs for the Darwin Tree of Life Project
AUTHORS: Jordan Beasley, Rebekka Uhl, Laura L Forrest, David Bell, Michelle Hart, Rebecca Yahr, Estelle Kilias

[DESCRIPTION]
This collection of protocols describe the standard operating procedures for DNA barcoding in the Darwin Tree of Life project.
The SOPs cover barcoding of macroalgae and marine protists, fungi and lichens at the Marine Biological Association, protists at the University of Oxford, plants and lichens at the Royal Botanic Gardens of Edinburgh and animals at the Natural History Museum of London.


[STEPS]
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p>This protocol outlines the steps taken to support sexual reproduction in Aiptasia lab cultures. It also provides steps for handling embryos and larvae, and basic husbandry to keep them alive.</p>
<p>Spawning protocol based on the methods from Grawunder et al. 2015.</p>

[STEPS]
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TITLE: Single-step assembly of double guide plasmid (pCas9-Duo) for gene-editing in Plasmodium 
AUTHORS: Abhinay Ramaprasad

[DESCRIPTION]
This protocol describes a one-pot GoldenGate assembly of a new dual-guide targeting plasmid (pCas9-Duo) expressing two distinct guide RNAs in order to enhance the chances of a successful Cas9-mediated modification at a target region in Plasmodium falciparum. Designed as part of SHIFTiKO (frameshift-based trackable inducible knockout) system1 (based on2).

[STEPS]
SECTION: gRNA oligo design
1. Add the overhangs "ATTG" and "AAAC" to forward and reverse oligos of gRNA1, and "TTGG" and "TAAA" to gRNA2 respectively.
 

SECTION: Anneal gRNA oligos
2. Set up annealing reactions. 
 
1.0 µL  gRNA.F 100 micromolar (µM) 
1.0 µL  gRNA.R 100 micromolar (µM) 
1.0 µL    
0.5 µL     
6.5 µL   
SECTION: Anneal gRNA oligos
3. Step 1  37 °C  30 min 
Step 2  94 °C  5 min 
Step 3  90 °C to 25 °C  RAMP 5 °C
Step 4  4 °C  Hold
SECTION: Single-step GoldenGate assembly
4. Make 1:200 dilution mixture of annealed gRNA1 and gRNA2.

1 µL gRNA1 annealed reaction
1 µL gRNA2 annealed reaction
198 µL   
SECTION: Single-step GoldenGate assembly
5. Set up assembly reaction

2 µL  pCas9 200 ng/µL 
1 µL  1:200 diluted oligo
0.5 µL    
0.5 µL    
2 µL    
2 µL   
12 µL   
SECTION: Single-step GoldenGate assembly
6. Step 1  37 °C  5 min 
Step 2  16 °C  5 min 
Repeat Step 1-2 for 6 cycles
Step 3  4 °C  Hold
SECTION: Transform, plate and screen colonies
7. Add 2 µL  of assembly reaction to 15 µL ultracompetent cells (like   ). 
SECTION: Transform, plate and screen colonies
8. Place on ice  for 20 min . 
SECTION: Transform, plate and screen colonies
9. Heat shock at 42 °C for 40 s  and spread transformed cells on  .
SECTION: Transform, plate and screen colonies
10. Pick colonies, miniprep-isolate plasmids and screen for gRNA1 and gRNA2 insertions by Sanger sequencing using   (CAGGAAACAGCTATGAC) and   (ACTGGCCGTCGTTTTAC), respectively.
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p><img id="s-mce-img" class="s-mce-img" src="https://s3.amazonaws.com/pr-journal/j4gdmie.png" data-src="https://s3.amazonaws.com/pr-journal/j4fdmie.png" data-ofn="cpib_root_analysis.png" /></p>

[STEPS]
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TITLE: Flow Cytometer Fluorescence Voltration for FCMPASS
AUTHORS: Joshua A Welsh, Sean M Cook, Jennifer  Jones

[DESCRIPTION]
Protocol to perform flow cytometer voltration to identify optimal detector settings for small particle analysis. Data acquired from this protocol are compatible with semi-automated analysis tools built into FCMPASS software.

[STEPS]
SECTION: Sample preparation
1. Vortex   bottle on a high setting for 5 s. 

SECTION: Sample preparation
2. Pipette 500 µL of   to two   . Label one tube 'DPBS" and the second tube 'Beads'.

 

SECTION: Sample preparation
3. Add 3 drops of   to the 'Beads'   and vortex for r 5 s. 

SECTION: Cytometer Setup
4. Ensure cytometer is clean and that -Height and -Area statistics are set to be collected on all parameters and that all parameters are on.
SECTION: Cytometer Setup
5. Create a pseudocolor plot with FSC-A on the X-Axis and (488 nm) B-SSC-A on the Y-Axis and make sure both parameters are being plotted on a linear-scale.
SECTION: Cytometer Setup
6. Create a histogram plot with (405 nm) V-SSC-H on the X-Axis and make sure it is plotted on a log-scale.
SECTION: Cytometer Setup
7. Set the cytometer triggering threshold to (405 nm) V-SSC-H. All samples should be acquired with the lowest flow rate, typically ~10-15 µL min-1.
 

SECTION: Cytometer Setup
8. Acquire the DPBS tube while viewing the histogram plot from  . Adjust the detector gain or trigger threshold until the instrument noise is being acquired at ~1000 events/sec. The instrument noise floor is distinct from detected background events in sheath as it has a sharp increase. In a system with debris there may be a tail that elongates out of this this sharp peak. 
  

SECTION: Cytometer Setup
8.1. Recording this noise is not necessary as this step is identifying optimal settings.
SECTION: Cytometer Setup
9. Acquire the 'Beads' tube from   . Using the plot from    adjust the FSC and B-SSC gain until the single bead population is clearly visible and can be easily gated from the doublet population to the top right of it. 
 
  

SECTION: Cytometer Setup
10. Creating a gate around the single bead population named 'Gate 1';
SECTION: Cytometer Setup
11. Adjust the stopping criteria of the instrument to record until 10,000 events are acquired on 'Gate 1' drawn in   .

SECTION: Performing Voltration
12. Voltration can now be performed by recording the 'Beads' tube at multiple fluorescent detector gains, leaving the trigger threshold and light scatter gains consistent. It is recommended that a recording of at least 10 fluorescent detector settings is taken. Including more increments within a voltration will result in being more confident of the subsequent optimal detector settings.  
 

SECTION: Performing Voltration
12.1. To ensure accurate data analysis, the brightest bead must be visible on at least two of the selected gains.
TITLE: Quantitative PCR of Viral Abundance
AUTHORS: Bryan  Yoo, Jessica  Griffiths, Sarkis  Mazmanian

[DESCRIPTION]
Protocol for quantifying viral abundance by PCR used in Yoo et al 2021

[STEPS]
1. AAV-PHP.S:hSYN1-mNeonGreen was delivered systemically to wildtype mice at 6-8 weeks of age.
2. 3-4 weeks following viral infection, 1cm of proximal, medial, and distal small intestine (SI) and colon were harvested and flash frozen in TRIzol (ThermoFisher Scientific, Waltham, MA-Cat. No. 15596018) and RNA was extracted per manufacturer’s instructions.
3.  Viral ssDNA is present in the RNA fraction. Contaminating RNA was eliminated with RNase treatment (ThermoFisher Scientific, Waltham, MA-Cat. No. AM228) per manufacturer’s instructions.
4. Quantitative PCR was performed on viral DNA with primers against WPRE (Forward: 5’- GGCTGTTGGGCACTGACAAT-3’; Reverse: 5’-CCGAAGGGACGTAGCAGAAG-3’) and values were normalized to ubiquitous mitochondrial gene, mtRNR2 (Forward: 5’- CCGCAAGGGAAAGATGAAAA-3’; Reverse: 5’-TCGTTTGGTTTCGGGGTTTC-3’). 
5. Plotted relative abundances are mean values across three regions sampled in either SI or colon.
TITLE: Distribution of aerophilous diatom communities associated with terrestrial green macroalgae in the South Shetland Islands, Maritime  Antarctica - PROTOCOL
AUTHORS: Juliana Ferreira da Silva, Maria Angélica Oliveira, Raylane Ribeiro Anunciação, Eduardo Pereira da Silva, Rodrigo Paidano Alves, Adriano Luis Schunemann, Filipe de Carvalho Victoria, Margéli Pereira de Albuquerque, Antonio Batista Pereira

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><span>The establishment of diatom communities depends on environmental factors such as the type of substrate and geographic conditions that influence the dispersal processes of these organisms. The main goal of this study was to evaluate the similarity between diatom communities associated with the macroalgae </span><span style = "font-style:italic;">Prasiola crispa</span><span>(Lightfoot) Kützing in relation to spatial distance from six sampled sites located in the South Shetland Islands, Maritime Antarctica. The diatom flora associated with </span><span style = "font-style:italic;">Prasiola crispa</span><span>was represented by 23 species distributed in 15 genera.  </span><span style = "font-style:italic;">Pinnularia australoschoenfelderi</span><span>Zidarova, Kopalová & Van de Vijver</span><span style = "font-style:italic;">, Luticolaaustroatlantica</span><span>Van de Vijver, Kopalová, S.A.Spaulding & Esposito, </span><span style = "font-style:italic;">Luticolaamoena</span><span> Van der Vijver, Kopalová, Zidarova & Levkov, </span><span style = "font-style:italic;">Pinnularia austroshetlandica</span><span>(Carlson) Cleve-Euler and </span><span style = "font-style:italic;">Psammothidium papilio</span><span>(D.E. Kellogg et al.) Kopalová & Zidarova were the most abundant species in our samples, which together represented 68% of the total number of individuals collected. There is a great similarity in composition and abundance of the diatom community among the sampled points., which resulted in the absence of a linear pattern of relationship with the distance between sampling points. We conclude that distance was not a factor of differentiation of Antarctic diatom communities associated with terrestrial green macroalgae. Suggesting that Antarctic environments may have unique characteristics with homogeneous abiotic factors, at least related to this substrate.</span></div></div>

[STEPS]
?.
TITLE: Western blot analysis and immunoprecipitation assay
AUTHORS: Marzia Ognibene

[STEPS]
?. Cells are washed in cold PBS and lysed on ice in a buffer containing 1.6 mM NaH2PO4, 8.6 mM Na2HPO4, 1% Triton X-100, 0.1% SDS; 0.1% NaCl, 0.5% NaDoc, 2 mM AEBSF, 20 mg/mL each of aprotinin and leupeptin.
?. Cell lysates are centrifuged in a microfuge at maximum speed for 10 min to eliminate debris.
?. Protein content is evaluated by bicinchoninic acid assay (BCA Protein assay kit by Thermo Fisher) in a spectrophotometer.
?. Lysates (100 µg each) are subjected to SDS-PAGE electrophoresis and transferred to PVDF membrane.
?. For the IMMUNOPRECIPITATION ASSAY, cells are lysed in a buffer containing 20 mM Tris-HCl, 1% Triton X-100, 10% Glycerol, 2 mM AEBSF, 20 mg/mL each of aprotinin and leupeptin.
?. Lysates (1000 µg) are incubated overnight at 4˚C with the desired antibody under constant rotation.
?. Gammabind G-Sepharose (40 µl) is added to each sample and let rotate for 1 hour and 30 min.
?. Samples are washed three times with the lysis buffer in a microfuge at 4300 g for 1 min.
?. After eliminating the last supernatant, samples are eluted in 40 µl of 1x Laemmli’s buffer, subjected to SDS-PAGE and transferred to PVDF membrane.
?. PVDF membrane is incubated in blocking buffer (4% nonfat milk in T-TBS -Tris Buffer Solution 1x with 0,05%Tween) for 2 hr at room temperature.
?. Membrane is incubated in primary antibody diluted in 0,5%BSA in T-TBS, overnight at 4°C.
?. Membrane is washed in T-TBS, 5 times, 5 min each.
?. Membrane is incubated in HRP-conjugated secondary antibody diluted in 0,5%BSA in T-TBS, 2 hr, at room temperature.
?. Membrane is washed in T-TBS, 5 times, 5 min each.
?. Proteins are visualized by enhanced chemiluminescence (ECL) detection, through the Bio-Rad ChemiDoc instrument. Signal intensity was measured by densitometry using the Bio-Rad Image Lab 6.0 software.
TITLE: Removal of gDNA from totalRNA
AUTHORS: iGEM Dusseldorf

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Removal of genomic DNA from RNA preparations</div></div>

[STEPS]
?. [Digestion of DNA]
pipet components into a RNase-free 1.5 ml tube (following order: H2O, buffer, total RNA, DNaseI)
?. [Digestion of DNA]
incubate reaction at 37°C
?. [Extraction of DNaseI-digested RNA]
add 1 µl 50 mM EDTA and incubate at 65°C
?. [Extraction of DNaseI-digested RNA]
add 1/10 Volume NaOAc and 3 Volumes 100% EtOH
?. [Extraction of DNaseI-digested RNA]
freeze at -20°C over night or at -80°C for at least 1h
?. [Extraction of DNaseI-digested RNA]
centrifuge at 13000 rpm at 4°C
?. [Extraction of DNaseI-digested RNA]
Discard supernatant RNA pellet may be invisible, make sure to align tube lid so you remember where the pellet is located
?. [Extraction of DNaseI-digested RNA]
add 500 µl 70% EtOH
?. [Extraction of DNaseI-digested RNA]
centrifuge at 13000 rpm at 4°C
?. [Extraction of DNaseI-digested RNA]
discard supernatant, remove as much ethanol as possible
?. [Extraction of DNaseI-digested RNA]
dry tubes at RT under the hood (~5min), do not overdry!
?. [Extraction of DNaseI-digested RNA]
resuspend pellet in 20 µl RNase-free water
?. [Extraction of DNaseI-digested RNA]
measure concentration
?. [Quality control:]
perform PCR on your DNA-free RNA. Include a positive control using gDNA as a template and primer that bind to it.There should be no visible bands except for positive control!
?. [Digestion of DNA]
Calculate the volume required for 1 µg RNA  AB1Componentamount2DNaseI buffer1 µl3total RNA1 µg4DNaseI1 µl5H2Oto 10 µl
AB1Componentamount2DNaseI buffer1 µl3total RNA1 µg4DNaseI1 µl5H2Oto 10 µl
TITLE: No Title
AUTHORS: 

[GUIDELINES]
<p>Overall Notes:</p>
<ul>
<li>Measure all regions twice (on separate days)
<ul>
<li>Use Pearson coefficient to determine how similar they are</li>
<li>If Pearson coefficient is 0.9, then average them together</li>
<li>If not, measure regions again and re-calculate the Pearson’s coefficient to each old sample</li>
<li>Average the samples that have Pearson’s coefficient of 0.9</li>
</ul>
</li>
<li>Measure all samples for given region on the same day
<ul>
<li>Light will change readings, will never be the same day-to-day</li>
</ul>
</li>
<li>Close the door to the scope room to help control for lighting differences</li>
<li>Lightbox is fluorescent, so as it warms up it will change lumens
<ul>
<li>let it warm up fully before starting anything</li>
</ul>
</li>
<li>If a region is done during the morning, and you are going to complete another region during the afternoon, leave everything on while you eat luch</li>
</ul>
<p>Cover the lightbox with the blue lab bench sheet</p>

[STEPS]
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TITLE: Open-source tools for Spatial Tissue Exploration Program (STEP) PhenoCycler-Fusion image analysis.
AUTHORS: Daniel H Geschwind, George Chen

[DESCRIPTION]
Link to PhenoCycler User Guide to allow for multi-plex spatial proteomic detection on fresh frozen and FFPE tissue sections plus image analysis. 

[STEPS]
SECTION: PhenoCycler Imaging Analysis protocol
2.   
Imaging Analysis protocol can be found here:
This protocol is working for us.
SECTION: STEP protocol for PhenoCycler coverslip preparation
1. STEP protocol for PhenoCycler coverslip preparation can be found here:  
This protocol is working for us.
TITLE: Hamster infestation with cercariae
AUTHORS: Frédéric D. Chevalier

[DESCRIPTION]
This protocol describes the steps, materials and precautions needed for infecting hamsters with schistosome cercariae. Note that the use of vertebrates for research needs to be approved by the Institutional Animal Care and Use Committee (IACUC). Refer to your local IACUC for more information.

[STEPS]
1. Shed cercariae from snails during 2h under light in TBRI water
2. Discard the shedding water in a new beaker
3. Shed cercariae from snails again during 2h under light in new TBRI water
4. Transfer the water in a new dish
5. Get the hamster
6. Wet the hamster in jar containing hot water (~30°C). BE CAREFUL to keep always the same individuals together (no mix from different cages)
7. Take 1,000 cercariae with thinned pipette and depose them in the pot will receive the hamster
8. Add water in the pot if necessary
9. Put the hamster in the pot and label the pot correctly (note the number of the cage and the strain used for the infestation)
10. Wait 2h minimum
11. Put the hamster in the cage. BE CAREFUL to keep always the same individuals together (no mix from different cages)
12. Wait until the hamsters were dried and go back to the animal room
13. Discard the biohazard materials in appropriate containers
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
Immunocytochemistry for morphology analysis. Staining of cell culture in order to obtain fluorescence images of the culture.

[STEPS]
SECTION: Cell fix
?. 
SECTION: Staining
?. 
SECTION: Permeabilization
?. 
SECTION: Block
?. 
SECTION: Primary antibody incubation
?. 
SECTION: Secondary antibody incubation
?. 
SECTION: DAPI stain
?.
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p>Testing antibacterial activity of bacteriophages on hard surfaces</p>

[BEFORE_START]
<p> </p>
<p>The following materials are needed:</p>
<p>- individual pure bacterial cultures</p>
<p>- bacteriophage preparation (either commercial or home-prepared)</p>
<p>- bacterial growth media (liquid and agarized)</p>
<p>- Rodac contact agar plates</p>
<p> </p>

[GUIDELINES]
<p>To test the killing/decontaminating activity of bacteriophages against specific bacterial targets.</p>

[STEPS]
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
The following procedure is an adaptation of the original FLVP protocol from Hennes et al. (1995) for use with SYBR Green I stain. Normally, this protocol will take 2 days given that the softening of the viral pellet is an overnight step. However, if the viral pellet redissolves quickly, it can be accomplished in 1 day.

[GUIDELINES]
The virus–host system used for the FLVP experiments was <em>Vibrio alginolyticus</em> strain PWH3a, a marine heterotrophic bacterium, and its phage PWH3a-P1, a species-specific dsDNA virus of the <em>Myoviridae</em> family. Both strains were originally isolated from the coastal waters of the Gulf of Mexico (Suttle and Chen 1992). The host was maintained as a –80°C glycerol stock to minimize any long-term culturing effects and was cultivated at 30°C with agitation (120–200 rpm) using Marine Luria-Bertani broth (MLB) (0.5 g L<sup>–1</sup> each of casamino acids, peptone, and yeast extract, 0.3% vol/vol glycerol, in 25 psu ultrafiltrate base). The virus was amplified using the plate lysate/liquid elution method (Suttle 1993), substituting ultrafiltered (virus-free) seawater for sterile media as the eluting agent. The eluant from multiple plates was pooled into a 50-mL centrifuge tube and spun at ~4000g for 20 min to remove large debris. The supernatant was then collected and filtered through a 47-mm–diameter, 0.22-µm–pore-size Durapore (Millipore) membrane to remove any remaining host cells. The final viral stock was titered by plaque assay (Suttle 1993) and kept at 4°C in the dark until needed.<br /><br /><strong>Materials and Equipment:</strong> <br /><br /> • amplified, concentrated virus stock (preferably ≥10<sup>10</sup> viruses mL<sup>–1</sup>); calculate amount of stock needed using the following guide: 1 tube (1.7 mL) of stock = 50 µL of FLVPs → 1 µL FLVPs/slide = 50 slides<br />• Screw cap 1.7-mL microcentrifuge tubes (often listed as 1.5 mL) <br />• 0.02-µm filtered water or appropriate filtered seawater medium for resuspensions; use the latter if the virus is destroyed by freshwater <br />• SYBR Green I dye working stock (Molecular Probes) <br />• RC80 Beckmann (or similar) ultracentrifuge with an SW40 (or similar) swinging-bucket rotor; alternatively, use a rotor capable of directly accepting microcentrifuge tubes in a centrifuge capable of reaching &gt;50,000g  <br />• epifluorescence microscope equipped with a 100× oilimmersion objective and a blue-light excitation filter (such as the Olympus U-MWB2/U-MWIB2 filters)

[STEPS]
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TITLE: Deepwell Reusual Protocol 
AUTHORS: Morris Jack

[DESCRIPTION]
This is a protocol to reuse deepwells for sustainability practices

[STEPS]
SECTION: Growing cultures in a deep well
1. Cover a deep well with two layers of aluminium foil and tape the front and back to hold it securely. The tape also turns the aluminium foil into a lid for the deep well by temporarily peeling off the front tape.
SECTION: Growing cultures in a deep well
2. Autoclave at 120C for 20min. After autoclaving, keep in a heated oven (> 50C) overnight for drying and desiccation.
SECTION: Growing cultures in a deep well
3. Do not fill the deep wells more than 60% of their max volume if they are to be placed in a shaking incubator to avoid spillage.
SECTION: Deep well Decontamination
4. Tape around the deep well over the aluminium foil to prevent spillage.
SECTION: Deep well Decontamination
5. While carrying the deep well outside the lab space, enclose it in a waterproof autoclave bag. Keep sufficient absorbent paper on hand in case of accidental spillage.
SECTION: Deep well Decontamination
6. In the autoclave facility, autoclave the deep wells at 120C for >30min to decontaminate.
SECTION: Deep well Decontamination
7. Once autoclaved, the deep wells will be brought back to the lab
SECTION: Deep well Decontamination
8. Carefully peel the aluminium foil and dispose the cultures and aluminium foil into a disposal bag containing paper roll to absorb the liquid.
SECTION: Deep well Decontamination
9. Send decontaminated culture for incineration (yellow bin route).
SECTION: Deep well Decontamination
10. Rinse the deep well plate with water and a brush to clean before future use.
SECTION: Deep well Decontamination
11. Keep in a heated oven (> 50C) overnight for drying and desiccation