eamag/protocols-with-steps huggingface dataset:
TITLE: Overnight Bacterial Culture
AUTHORS: Alba Balletbó

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">This protocol describes how to grow an overnight culture of bacteria in LB (Luria-Bertani) liquid media broth. </div></div>

[STEPS]
?. Pour appropriate volume of LB broth into the selected autoclaved container.
?. Open and add the frozen aliquot of bacteria to the LB broth or making use of an inoculation loop, carefully select your isolated colony from the media plate and shake the loop in media until the fragment has come loose and is visibly floating in the media.
?. Loosely seal the container, either using tape to secure the loosely fitted lid of a tube or tinfoil to cover a flask.
?. Place the tube/flask in an orbital shaker at the optimal growing temperature of your bacterial species (this will likely be 30 or 37° C). Leave the liquid culture to grow overnight.
TITLE: P1 Kidney Cold-Active Protease Single Cell Dissociation
AUTHORS: Andrew Potter, Steve  Potter

[DESCRIPTION]
Method used to derive single cell suspension from P1 mouse kidneys on ice, generating a cell suspension with greatly reduced artifact gene expresion changes and suitable for downstream analysis using 10x Chromium or DropSeq. 

[GUIDELINES]
 Storage Conditions of Reagents 
  
 
 
  
  
  Required Equipment
  
 
 
  
  
 The protocol workflow is as follows: 
  
    A. Isolate Kidney
    B. Initial digestion: triturate on ice
    C. Perform gentleMACS 
    D. Continue triturating on ice
    F. Preparing cells for Chromium/DropSeq
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
  BEFORE STARTING
   Prepare Bacillus Licheniformis enzyme mix just prior to starting dissociation:
  
 
   Volume (µl) Reagent  Final concentration     894  DPBS  1X    1  0.5 M EDTA  0.5 mM    5  DNAse 1 (250 U/10 μL)  125 U / mL    100  B. Lich (100 mg/mL)  10 mg/mL   
    +25 mg tissue / 1 mL enzyme mix
 
 
 
 
 
 
 
 
To prepare 0.01% BSA/PBS:
 
Make stock of 10% BSA in DPBS and store at -20 °C. To make PBS/BSA 0.01% aliquot 50 mL of DPBS in 50 mL conical and pipet in 50 µL of 10% BSA stock.
 
Prepare 10% FBS/PBS with heat-inactivated FBS.
 
 

[STEPS]
1. Extract & isolate P1 kidneys in ice-cold PBS.
2. Mince kidneys on top of petri dish, on ice, using razor blade.
3. Weigh out 25 mg of tissue for each tube of B. Lich. enzyme mix (2 tubes total). 

25 5
4. Incubate tissue + enzyme on ice for 7 minutes while triturating 15 strokes using 1 mL pipet every 2 minutes set to 700 µL - first with tip cut off.

7 min
2 min
5. After 7 minutes, take the digest mix (combine the two tubes) and pipet into Miltenyi C-tube (placed on ice); take C-tube to gentleMACS placed in 4° cold room. Run program brain_03 two times. 

4 10
6. After MACS, briefly quick spin the MACS tube (to 300 G) at 4 °C to ensure contents are in the bottom of the tube. 

4 10
7. Re-suspend and visualize cells using scope by taking small aliquot and using a slide; continue digesting cells in C-tube on ice for 8 additional minutes while triturating every 2 min 15 strokes using a 1 mL pipet.

8 min
2 min
8. Add 3 mL ice-cold 10% FBS/PBS to digest mix in C-tube to inhibit the protease. 

3 2
9. Transfer digest mix to a 15 mL conical. Spin 300 G for 5 minutes at 4 °C; discard supernatant; re-suspend cell pellet in 2 mL ice-cold PBS/BSA.

4 10
5 min
2 2
10. Filter re-suspended cells using 30 uM filter on sterile 15 mL conical on ice - rinse filter with 8 mL ice-cold PBS/BSA.
 
8 2
11. Spin 15 mL conical tube containing filtered cells 300 G for 5 minutes at 4 °C; discard supernatant and re-suspend pellet in 10 mL ice-cold PBS/BSA.

4 10
5 min
10 2
12. Repeat rinse/spin in ice-cold PBS/BSA.
13. Remove supernatant and re-suspend in 1-2 mL ice-cold PBS/BSA.
14. Examine using hemocytometer and adjust concentration to 100 cells/uL for DropSeq or 1,000 cells/µL for 10X Chromium.
TITLE: Immunoprecipitation of NAP1-GFP
AUTHORS: Elias Adriaenssens

[DESCRIPTION]
This protocol describes immunoprecipitation of NAP1-GFP from HAP1 cells.

[STEPS]
SECTION: Immunoprecipitation
1. Transiently transfect FIP200 knockout HAP1 cells with pcDNA3.1 NAP1-EGFP (Addgene), or empty pcDNA3.1 vector as a negative control, using Lipofectamine 2000 (Thermo Fisher).
 

SECTION: Immunoprecipitation
2. After 48 h, collect the cells by trypsinization and wash the cell pellet with PBS once. Then, lyse the cells in lysis buffer.
SECTION: Immunoprecipitation
3. Lyse the samples for 20 min on ice. Then, clear the cell lysates by centrifugation at 20000 x g, 10 min, 4 °C.

SECTION: Immunoprecipitation
4. Determine the protein concentrations of the cleared protein lysates with the Pierce Detergent Compatible Bradford Assay Kit (23246, Thermo Fisher).
SECTION: Immunoprecipitation
5. For both samples, negative control and NAP1-EGFP lysates, incubate 12 mg of cell lysate  20 min with 20 µL or GFP-Trap agarose beads (GTA-20, Chromotek).
SECTION: Immunoprecipitation
6. In the morning, wash the samples three times in lysis buffer. Then, resuspend the beads in protein loading dye, supplemented with 100 mM DTT. Finally, boil the samples for 5 min at 95 °C.

SECTION: Immunoprecipitation
7. Load the samples on 4-12% SDS-PAGE gels (NP0322BOX, Thermo Fisher) with PageRuler Prestained protein marker (Thermo Fisher). 
SECTION: Immunoprecipitation
8. After the run, stain the SDS-PAGE gel with Coomassie, fix for 10 min in fixation solution, and destain it 5 min in dH2O.

SECTION: Immunoprecipitation
9. Cut the band corresponding to NAP1-EGFP from the gel with a fresh scalpel and submit for mass spectrometry analysis.
TITLE: Von Stosch (VS) enriched seawater medium solution
AUTHORS: Anton Kuech

[DESCRIPTION]
This protocol outlines the steps required to prepare von Stosch (VS) enriched seawater medium modified according to Guiry & Cunningham (1984).
 


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

[STEPS]
SECTION: Vitamin B12 stock solution
1. 5 mg   in 500 mL  
 

SECTION: Primary solution
1.1. In 4 L   dissolve:
SECTION: Primary solution
1.2. 1.86 g   (Na₂-EDTA 2 H₂O)
SECTION: Primary solution
1.3. 21.26 g   
SECTION: Primary solution
1.4. 0.139 g   
SECTION: Primary solution
1.5. 0.98 g   
SECTION: Primary solution
1.6. 0.05 g   
SECTION: Primary solution
1.7. 0.1 g   
SECTION: Primary solution
1.8. 20 mL  Vitamin B12 stock solution (5 mg in 500 mL    )
SECTION: Primary solution
1.9. 10 mg   
SECTION: Primary solution
1.10. Pasteurise for 120 min  at 99 °C 
SECTION: Storage
2. Store VS solution in Duran bottles in a cold and dark setting at 10 °C .
 

SECTION: Application
3. Culturing medium is created by adding 200 mL VS to 10 L sterile seawater
 

None. 

5 mg   in 500 mL
TITLE: Subsampling Ethanol Preservative from Zooplankton Museum Collections for DNA Extractions
AUTHORS: Andreas Novotny

[DESCRIPTION]
This protocol is used to sample DNA from archived samples preserved in ethanol, without having to subsample or split the actual zooplankton biomass.

[BEFORE_START]
Read background information, MIOP and BePOP-OBON information under the "Guidelines" tab.

[GUIDELINES]
 MIOP: Minimum Information about an Omics Protocol
 
 
 
 
AUTHORS
 
 
 

RELATED PROTOCOLS
 
 
ACRONYMS AND ABBREVIATIONS
 
 
GLOSSARY
 
 

BACKGROUND
This protocol is used to sample DNA from archived samples preserved in ethanol, without having to subsample or split the actual zooplankton biomass.

Spatial coverage and environments of relevance
As part of the Hakai Institute Ocean Observing Program, biomolecular samples have been collected weekly, from 0 to near bottom (260 m), to genetically characterize plankton communities in the Northern Salish Sea since 2015, developing a climatology from which we can begin uncover the physical, chemical and biological drivers of community and functional change in the dynamic coastal waters of coastal British Columbia. 

This protocol has been used as an alternative source of genetic information, when it is not practical to remove biomass from the zooplankton samples.

Personnel Required
 1 Technician

Safety
Identify hazards associated with the procedure and specify protective equipment and safety training required to safely execute the procedure!

Training requirements
Sterile technique, pipetting skills. Work-safe laboratory practices.

Time needed to execute the procedure
1 h

[STEPS]
SECTION: ETHANOL SUBSAMPLING
2. To re suspend DNA in the zooplankton sample, invert the sample jar three times.
SECTION: ETHANOL SUBSAMPLING
3. Let settle for 30 minutes
SECTION: ETHANOL SUBSAMPLING
4. Use a serological pipette to remove 50 ml of the ethanol preservative, and transfer to a 50 mL falcon tube.
SECTION: ETHANOL SUBSAMPLING
5. Use a syringe to push the ethanol through a   .

SECTION: ETHANOL SUBSAMPLING
6. Seal the outflow of the filter with parafilm.
SECTION: ETHANOL SUBSAMPLING
7. Add 1800 μL sucrose lysis buffer (SLB).
SECTION: ETHANOL SUBSAMPLING
8. Seal the inflow opening with parafilm.
SECTION: ETHANOL SUBSAMPLING
9. Label filter units and store them at -80°C for downstream DNA extraction.
SECTION: PREPARATIONS
1. Prepare Sucrose Lysis Buffer (SLB):
 

SECTION: DNA EXTRACTION
10. Follow the same extraction procedures as for Environmental DNA: 
 

SECTION: DNA EXTRACTION
11. Alternative extraction method:
TITLE: Sample preparation and lysis of homogenized malaise trap samples
AUTHORS: Dominik Buchner

[DESCRIPTION]
This protocol describes the steps of sample preparation and lysis before DNA extraction for the Malaise trap metabarcoding protocol of the LeeseLab.


[BEFORE_START]
Make sure all buffers are prepared before starting. 

[GUIDELINES]
Follow general lab etiquette. Wear gloves to prevent contaminating the samples. Clean the workspace before starting with 80% EtOH. 

[STEPS]
2. Shake the sample well.
Transfer 800 µL of the small size fraction and 200 µL of the large size fraction to a 2 mL screwcap tube. It might be beneficial to use wide-bore tips or cut off the tip when using regular pipette tips.

3. 11.000 x g, 3 min 

4. Remove as much ethanol as possible with a 1000 µL pipette. 

1. For each sample prepare a screwcap tube pre-filled with a few 2 mm zirconia beads.



5. Add 900 µL  of TNES buffer and 100 µL of Proteinase K working solution. Vortex shortly.

 

6. Bead-beat for 2 min at   

7. Incubate   

8. Store at -20 °C until DNA extraction.
TITLE: Neuropathy Phentoyping Protocols - Hematoxylin and Eosin Staining
AUTHORS: Eva Feldman

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Summary:</span></div><div class = "text-block"><span style = "font-weight:bold;">Phenotyping of Rodents for the Presence of Diabetic Neuropathy </span></div><div class = "text-block">In man, the development of diabetic neuropathy is dependent on both the degree of glycemic control and the duration of diabetes. Diabetic neuropathy is a progressive disorder, with signs and symptoms that parallel the loss of nerve fibers over time. Consequently, assessments of neuropathy in mice are not performed at one time point, but are characterized at multiple time points during a 6 month period of diabetes. The degree of diabetes is evaluated in 2 ways: tail blood glucose measured following a 6 hour fast and glycated hemoglobin levels. The initial degree of neuropathy is screened using the methods discussed below. Detailed measures of neuropathy are employed when the initial screening instruments indicate a profound or unique phenotypic difference. This document contains protocols used by the DiaComp staff to examine and measure diabetic neuropathy at the whole animal, tissue and cellular levels.</div><div class = "text-block"><span style = "font-weight:bold;">Diabetic Complication:</span></div><div style = "text-align :; float : ;"><img style = "" src = "https://s3.amazonaws.com/pr-journal/binb4kcn.jpg" /></div><div class = "text-block"><span style = "font-weight:bold;">Reference: </span></div><div class = "text-block"><span style = "font-style:italic;">AFIP Laboratory Methods in Histotechnology, 1994, p. 16, 53-55. </span></div></div>

[STEPS]
?. [H&E Procedure:]
For paraffin sections and fixed cryosections, deparaffinize and hydrate to water.
?. [H&E Procedure:]
For fresh cryosections, place in alcoholic formalin for 45 seconds and rinse in water.
?. [H&E Procedure:]
Stain in Mayer’s hematoxylin for 15 min.
?. [H&E Procedure:]
Wash in lukewarm running water for10 minutes.
?. [H&E Procedure:]
Rinse in distilled water.
?. [H&E Procedure:]
95% EtOH for one minute, agitating.
?. [H&E Procedure:]
Counterstain in eosin-phloxine solution for 2 minutes.
?. [H&E Procedure:]
Dehydrate and clear.
?. [H&E Procedure:]
Coverslip and mount.
TITLE: Preparation of tissue culture plates for neural rosettes and neural progenitors
AUTHORS: Celeste Karch, Rita Martinez, Jacob Marsh

[STEPS]
?. Coat 3 wells of a 6-well plate with Poly-L-Ornithine (PLO) for  or overnight in humidified incubator.
1 ml
?. Aspirate off PLO and rinse 3 times with DMEM/F12.
?. Dilute laminin to 10 μg/mL final in cold DMEM/F12 and add 1 mL/well of solution to PLO coated plates.
?. Incubate for  or overnight in humidified incubator.
TITLE: U Mass - Non-esterified fatty acids
AUTHORS: Jason Kim

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Summary:</span><span style = "font-weight:bold;"> </span></div><div class = "text-block">
This experiment measures serum and plasma concentrations of non-esterified fatty acids using a 96-well kit. The experiment involves a coupled reaction to measure non-esterified fatty acids (NEFA) which ultimately forms a purple product that absorbs light at 550nm. This allows the concentration of NEFA to be determined from the optical density measured at 540~550nm. Serum fatty acids levels reflect systemic lipid metabolism, lipid digestion/absorption, and lipid clearance. Serum fatty acids levels are altered in obesity, insulin resistance, and type 2 diabetes.</div></div>

[STEPS]
?. Add 5 μl (or 1~10 μl) of serum or plasma sample to a well of Plate A.
?. Add dilution buffer to each well to reach a total sample volume of 50 μl.
?. Addition of 5 μl results in a 10x dilution of sample (5 μl of serum/plasma sample in 50 μl total sample volume).
?. Add 50 μl of each standard to empty wells. Use Plate B if necessary.
?. Add 10.5 ml of the reconstituted FFA Reagent A to one of the disposable trays provided with the kit.
?. Add 100 μl of FFA Reagent A to each well.
?. Gently shake the plate to ensure thorough mixing.
?. Place in a 37ºC incubator for 10 minutes.
?. Add 5.5 ml of the reconstituted FFA Reagent B to the other disposable tray provided with the kit.
?. Add 50 μl of FFA Reagent B to each well.
?. Gently shake the plate to ensure thorough mixing.
?. Place in a 37ºC incubator for 10 minutes.
?. Allow the plate to equilibrate to room temperature for 5 minutes. During this time, ensure that there are no bubbles in the solution mixture.
?. Use a large gauge needle or clean pipet tip to pop any bubbles as this will result in inaccurate absorbance readings.
?. Measure the optical density of each well at 540 nm.
TITLE: 跑膠
AUTHORS: Yin-Tse Huang,  Tina

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

[STEPS]
?. + 配成1倍的buffer
[TBE]
[DDW]
?. 大片的膠大約小片的膠大約Agarose 1% 大塊的膠 表示加 和 DDW
30 mL
20 µl
0.3 g
30 mL
?. 把配好的Gel 1%  放入微波爐加熱 ，每 一次
?. 將Gel到入製膠模型，若有氣泡，可以吸管尖挑開，之後架上齒梳，等
?. 輕輕將齒梳向上拉開（注意要慢慢拉開，否則要加DNA樣品的孔洞會破掉)
?. 將膠體連同膠模一起放入電泳槽中
?. 加入 在電泳槽中
[TBE buffer]
?. 混和滴在 parafilm 上
[DNA]
[dye]
?.
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p>This protocol provides step-by-step instructions for implementing a perl script to easily automate quality checking of raw tag sequences. This was specifically constructed for the numerous 18S diversity projects (which target the V4 hypervariable specific for evaluting protistan diversity).</p>
<p> </p>
<p>It is highly recommended that you follow along this Github repo: https://github.com/shu251/QC_steps_V4_tagsequencing</p>
<p> </p>

[BEFORE_START]
<p>Required programs:</p>
<ul>
<li>Database (preferrably with associated taxa list) to align sequences to. A version of the PR2 database formatted for QIIME can be found <a href="https://drive.google.com/drive/folders/0Bxzw_UrYS4IAaEQ0b0lMQ0ZiUkU?usp=sharing" target="_blank">here</a></li>
<li><a href="http://qiime.org/install/install.html" target="_blank">QIIME</a> - v.1.9.1 or higher. Note as of Jan 2018 QIIME1 will not be supported. Future iterations of this protocol will use QIIME2.</li>
<li>fastqjoin - this should install with QIIME. Alternatively you can use <a href="https://sco.h-its.org/exelixis/web/software/pear/doc.html#installing" target="_blank">PEAR</a> to merge PE sequences.</li>
<li><a href="http://www.usadellab.org/cms/?page=trimmomatic" target="_blank">Trimmomatic</a> - v.0.32</li>
<li><a href="https://github.com/torognes/vsearch" target="_blank">vsearch</a> - v1.11.1</li>
<li>Make sure you know your primer sequences and generate a fasta file: 'trimPE_V4primer.fasta'. In the below example I'm using <a href="http://onlinelibrary.wiley.com.libproxy1.usc.edu/doi/10.1111/j.1365-294X.2009.04480.x/full/" target="_blank">V4 Stoeck et al. 2010 primers</a></li>
</ul>

[GUIDELINES]
<p>Protocol describes quality checking process for raw fastq sequences. Includes some discussion for alternate approaches. End product is ready for OTU clustering. I've also included suggestions for OTU clustering at the end (specific for 18S data). Much of this pipeline can be applied for any tag sequencing analysis, but keep in mind this was specifically constructed to analyze single-celled microbial eukaryotic communities.</p>
<p> </p>
<p>This protocol (and my preferred method) is to perform as much quality checking on each individual sample before combining for downstream OTU clustering (or other analysis). This way, I can track any samples that may be not ideal for downstream analysis.</p>
<p> </p>
<p>Full github repo: https://github.com/shu251/QC_steps_V4_tagsequencing</p>

[STEPS]
?. 
?. 
?. 
?. 
?. 
?.
TITLE: Strategies for optimizing the isolation and expansion of sensitive patient-derived duodenoids, ileoids and colonoids .pdf
AUTHORS: Katlynn Bugda Gwilt, Jay R. Thiagarajah

[DESCRIPTION]
These protocols are an optimization of previous protocols and the original work described in Sato et al., 2009 which provided a foundation for the development of primary duodenoids, ileoids and colonoids from human samples. Here, we optimized methods specifically for use with samples from pediatric patients with genetic intestinal epithelial disorders. These protocols apply specifically to enteroids generated from endoscopic biopsies from patients with Congenital Diarrheas and Enteropathies (CoDE) or Very-Early-Onset Inflammatory Bowel Disease (VEO-IBD) or age-matched patients without any intestinal disease and include samples from the duodenum (duodenoids), ileum (ileoids), and colon (colonoids). The duodenoids, ileoids and colonoids from these patients exhibit a range of cellular phenotypes that potentially impact enteroid growth and maintenance including increased apoptosis, defects in proliferation, polarity and vesicular trafficking. These characteristics make it potentially challenging for long-term culture and expansion, limiting the availability of cells for functional experiments. Using a modified culture media, an expanded initial culture time after cell isolation from patient biopsies, and gentle passaging techniques, we were able to successfully culture and expand several patient lines over multiple passages and maintain the cultures for >5 years. The media conditions and protocols described here allow for reproducible phenotypes as well as scaling for larger functional studies on patient lines. These protocols also provide a useful starting point for further optimization for generating and culturing enteroids from patients with novel disease pathophysiology.

[BEFORE_START]
Dissolve all reagents to the appropriate concentration according to manufacturer’s protocols. A * indicates that there were no modifications to the reagent.

[STEPS]
TITLE: BAF_Protocol_007a Chloroform/Methanol Precipitation
AUTHORS: Nicholas Sherman

[DESCRIPTION]
This protocol extends BAF_Protocol_007 as an alternative to acetone precipitation (normal method). The chloroform/methanol precipitation is a better method if the sample is high in lipid or other hydrophobic small molecules. This method also produces a protein pellet and not the normal liquid interface and as such is easier.

[GUIDELINES]
This precipitation would be substituted in for the acetone precipitation normally used in BAF_Protocol_007.

[STEPS]
SECTION: Chloroform/Methanol PPT
1. 100 uL of protein extract of sample added to Eppendorf tube.
SECTION: Chloroform/Methanol PPT
2. 400 uL of Methanol was added, vortexed well.
SECTION: Chloroform/Methanol PPT
3. 200 uL of Chloroform was added, vortexed well.
SECTION: Chloroform/Methanol PPT
4. 300 uL of Water was added, vortexed well.
SECTION: Chloroform/Methanol PPT
5. Sample was centrifuged at 13,000 rpm for 2 minutes
SECTION: Chloroform/Methanol PPT
6. The upper layer was carefully removed (not disturbing the protein interface).
SECTION: Chloroform/Methanol PPT
7. 300 uL of Methanol was added, vortexed well.
SECTION: Chloroform/Methanol PPT
8. Samples were centrifuged at 13,000 rpm, for 2 minutes.
SECTION: Chloroform/Methanol PPT
9. Supernatants were discarded and the protein pellet was air dried. Resuspend in 100 mM Ammonium Bicarbonate in volume based on protein amount.
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p>This protocol allows transformation of two species of <em>Pseudo-nitzschia </em>through biolistic method.</p>
<p>With this method it is possible to insert one (transformation) or two (co-transformation) plasmids in the cells. </p>

[STEPS]
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?.
TITLE: Sistema de categorías para el estudio de la eficacia escolar
AUTHORS: Carla Ortiz-de-Villate, Javier Rodríguez-Santero, Juan Jesus Torres-Gordillo

[DESCRIPTION]
Sistema de categorías para el estudio de la eficacia escolar.  Se ha calculado su confiabilidad y concordancia entre investigadores en nuestra investigación con un Kalpha de Krippendorff alto (α=0.807).

[STEPS]
TITLE: UPitt TriState SenNet TMC Donor acceptance criteria 
AUTHORS: John Sembrat, Alison Morris, koenigshoffm, Oliver Eickelberg

[DESCRIPTION]
This document outlines the required criteria for donor inclusion of lung and heart specimens in the TriState SenNet TMC Biospecimen Core at the University of Pittsburgh, as part of the Cellular Senescence Network Program (SenNet). The study intends to be as inclusive as possible of race and ethnicity, dependent only on the consent for donor tissue for research. Disease known at the time of donation or later identified during the research will not necessarily exclude the sample from the research program.

[STEPS]
SECTION: Inclusion Criteria
1. Inclusion Criteria:  
Age: Adulthood -Young through middle age and senior years (approx. 18 to >95 years)
All sex, gender, race, ethnicity are eligible
Known state of health and diagnoses
SECTION: Exclusion Criteria
2. Exclusion Criteria: 
Unknown time of death
Prolonged Warm Ischemic Time (more than 2h)
Prolonged Cold Ischemic Time (more than 18h)
Known diseases including congenital anomaly or inborn error unsuitable for SenNet research
Infection with high-level organisms representing risk in processing and shipment
TITLE: NCEM Drop - Conventional (TM-014)
AUTHORS: sandra.crameri

[DESCRIPTION]
Standard routine conventional negative contrast EM.

[STEPS]
SECTION: HEADER
1. SAN:




SPEC No:




OPERATOR:



SECTION: Conventional
2. Adsorb 10 µL sample to grid 10 min , inspect to ensure sample does not dry out.

SECTION: Conventional
3. Drain excess sample from grid using filter paper, leave wet.
SECTION: Conventional
4. Stain  1 min 

5. Drain & dry using filter paper
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
This is a protocol for making a trace metal clean 0.5 mol L<sup>-1 </sup>EPPS (<span style="font-weight: 400;">N-(2-hydroxyethyl)piperazine-N-(3-propanesulfonic acid) solution for dissolved cobalt analyses.</span>

[STEPS]
?. 
?. 
?.