eamag/protocols-with-steps huggingface dataset:
TITLE: Preparation of aqueous extracts of plants
AUTHORS: Cecilia Mónica Rodríguez García, Leticia Peraza

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Aqueous extracts of plants are a simple, economical and eco-friendly alternative to use as a source of antifungal activity. Here is a protocol for extracting any type of plant material (leaves) with water using a simple blender to grind the dried leaves with water and then a series of centrifuges to eliminate the solid waste, finally a series of filtrations to have an aqueous extract sterile. This aqueous extract can be stored at 4 ° C and depending on the plant the activity can be maintained for a fixed period of time. After this process the aqueous extract is ready to be used in bioassays of antifungal activity.</div></div>

[STEPS]
?. [Aqueous extract preparation from leaves]
-Collection and preparation of leaves for dryingCollect fresh leaves from the fieldIn the lab wash them with tap waterRemove excess water with a kitchen centrifuge
?. [Aqueous extract preparation from leaves]
-Drying of leavesDistribute the leaves in cardboard boxesPut them at 50 °C in a stove for 2 -3 daysStore them in plastic bags until use
?. [Aqueous extract preparation from leaves]
1. Blend in a laboratory blender at the maximum speed
[dry leaves]
[every 30 sec stir with a spatula]
?. [Aqueous extract preparation from leaves]
4. Take the supernatant and pour it into a container, take 1800 μL in 2 mL eppendorf tubes, use the necessary tubes5. centrifuge
[at 20817 g, room temperature]
?. [Aqueous extract preparation from leaves]
3. Pour the mixture into 50 mL falcon tubes and centrifuge
[at 3220 g]
?. [Aqueous extract preparation from leaves]
2. Add
[sterile distilled water]
[mix everything]
?. [Aqueous extract preparation from leaves]
6. Take the supernatant of all the tubes and pour it into a container7. Filter the aqueous extract with the membrane of 0.65 μm contained in a swinnex8. Take the previous filtering and filter it with the membrane of 0.45 μm contained in a swinnex
?. [Aqueous extract preparation from leaves]
9. In a laminar flow hood, sterilize the aqueous extract by filtration using a membrane of 0.22 μm contained in a swinnex10. Store the aqueous extract  in a sterile 50 mL falcon tube in the dark at 4 ° C until its use.
TITLE: Multi-Patch Clamp Recording
AUTHORS: Allen Institute for Brain Science

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">This protocol describes the process to obtain multiple electrophysiological recordings from postnatal mouse and/or human brain slices.</div><div class = "text-block"><span style = "font-weight:bold;">Note</span><span>: Research reported in this publication was supported by the National Institute Of Mental Health of the National Institutes of Health under Award Number U19MH114830. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.</span></div></div>

[STEPS]
TITLE: Interpenetration-between-a-polymer-star-and-a-polymer-brush
AUTHORS: Mike Edwards

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">By means of the density functional theory framework I tackle the  long-standing problem of a polymer star interpenetrating with a polymer  brush at thermal equilibrium. Remarkably, the star is repelled to the  outside of the brush once it sucks into the brush. It turns out that  there could be a highly fluctuating region at the brush edge. The highly  fluctuating region would be responsible for discontinuous absoption  transitions by brushes. However, up to an small interpenetration length,  below which asphericity of the star is maintained, the star gets  collapsed by sucking more and more into the brush.</div></div>

[STEPS]
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p>We use the described methods for determining the internal cellular quota of thiamin and its<br />different phosphorylation states in marine algae, in the below case for the haptophyte Emiliania<br />huxleyi. The chemical assay used – the thiochrome assay - was developed in the 1930s but has<br />been modified over the decades to include instrumentation with improved detection (Backstrom et<br />al. 1995, Reddick et al. 2001). The assay is based on cyclization/oxidation of thiamin to<br />thiochrome under basic conditions in the presence of an oxidizing agent such as potassium<br />ferricyanide. Thiochrome is intensely fluorescent and easy to detect and quantify with high<br />sensitivity in an HPLC assay. This method has previously been used to determine cellular quotas<br />of thiamin and its different phosphorylation states in algae from brackish waters (Pinto et al. 2002,<br />Sylvander et al. 2013).</p>

[STEPS]
?. 
?. 
?. 
?. 
?.
TITLE: Freezing of mouse embryonic fibroblasts (MEFs) for hPSC cultures
AUTHORS: Hanqin  Li, Oriol Busquets, Steven Poser, Dirk Hockemeyer, Frank Soldner

[DESCRIPTION]
This protocol describes the freezing of mouse embryonic fibroblasts (MEFs), which can later be used as feeder cells for human pluripotent stem cell (hPSC) culture. 

General notes
1. This protocol can be used to freeze MEFs before or after inactivation.
2. Throughout this protocol, the term hPSC is used to collectively refer to both hiPSCs and hESCs. All described procedures have been tested and work equally well for hiPSCs and hESCs.
3. MEFs were obtained as described in Manipulating the Mouse Embryo: A Laboratory Manual, Third Edition (ISBN: 0879695919)
 




[STEPS]
1. Wash the plates twice with DPBS 
2. Add Trypsin (5 ml for 15-cm plate) and incubate for  5 min  (37°C; 5% CO2)

3. Add 10 ml MEF medium to neutralize the Trypsin and collect the solution into a conical tube.
4. Centrifuge the cell suspension at 250 x g, 10 min 


5. Discard supernatant, and re-suspend the cells in MEF Freezing Medium I at a concentration 2X the desired final freezing concentration (Freezing concentration will vary with MEF usage.)
6. While swirling, slowly and drop-wise, add an equal volume of MEF Freezing Medium II to the cell suspension.
7. Dispense cell mixture into pre-labeled cryovials (10x106) including type of cells, passage number, date frozen, and who handled cells.
8. Place cryovials into Styrofoam microtube freezer box or NALGENE™ Cryo 1°C Freezing Container pre-filled with 250 ml room temperature isopropanol.(This ensures that a -1°C/min rate of cooling is achieved, which is critical to cell viability.)
9. Freeze at   -80 °C 

10. For long term storage, store cryovials in liquid nitrogen (-196ºC).
6.1. MEF Freezing Medium II
 

3.1. MEF medium
 

5.1. MEF Freezing Medium I
TITLE: Inoculum preparation for Mixotrophic culturing
AUTHORS: Bruna Emerenciano, Maria Izabel Pereira Batista, Adriano Henrique Rangel

[STEPS]
?. Prepare the inoculum from Spirulina platensis strain to inoculate 800 mL of Zarrouk medium contained in 1000 mL Erlenmeyer flask.
?. Supplement the Zarrouk medium with 2.5%, 5.0% and 10% clarified buffalo mozzarella cheese whey. The components of Zarrouk medium (per liter) are as follows: 16.8g NaHCO3, 0.5g K2HPO4, 2.5g NaNO3, 1.0g K2SO4, 1.0g NaCl, 0.2g CaCl2, 0.04g MgSO4•7H2O, 0.08g EDTA,  1ml of A5 solution, 1ml of B6 solution.
?. Keep the inoculum growing mixotrophically for 4 days on experimental apparatus under the same light intensities as the culture inoculated by it.
?. Keep the cultures under temperature of 25 °C and constant sterile aeration promoted by pumps (JAD Air Pump S-510) at a specific flow rate of 0.5 vvm (volume of air per volume of medium per minute), and the only light source from fluorescent lamps of 45 W with a luminous intensity of 238 μmol m-2 s-1measured by Q201 quantum radiometer (Macam Photo-Metrics Ltd., Livingston, Scotland).
?. Use a light regime of 12-hour light, 12-hour darkness
TITLE: 2-step PCR mixture and conditions (Barcoded-head primers for seqs pooling)
AUTHORS: Yin-Tse  Huang

[DESCRIPTION]
PCR mixture and condition (2X SUPERGREEN PCR MASTER MIX)

[STEPS]
SECTION: For 1&#x27; PCR head-primers
2. Prepare 1' PCR master mixutre for head-primers (prepare 1.2X of solutions for pipetting error if needed)
 
PCR mixture for head-primers for each reaction
 

SECTION: For 1&#x27; PCR head-primers
7. Carry out PCR using the following condition:  

1' PCR condition for head-primers
 

SECTION: For 1&#x27; PCR head-primers
7.1. 1' hear-primers used in Huang lab
 

SECTION: For 2&#x27; PCR barcoded-head primers
16. Carry out 2' PCR using the following condition:  

2' PCR condition for barcoded-head primers
 

SECTION: For 2&#x27; PCR barcoded-head primers
16.1. 2' barcoded-head primers used in Huang lab
 

SECTION: For 2&#x27; PCR barcoded-head primers
10. Prepare 2' PCR master mixutre for barcoded-primers (prepare 1.2X of solutions for pipetting error if needed)
 
PCR mixture for barcoded-primers for each reaction (NO PRIMERs at this point!!)
 

SECTION: For 1&#x27; PCR head-primers
4. Transfer  15 µL 1' PCR master mixutre in 8-strip PCR tubes.

SECTION: For 2&#x27; PCR barcoded-head primers
12. Transfer 21.5 µL of the 2' PCR master mixture to PCR tubes.

SECTION: For 1&#x27; PCR head-primers
8. Carry out electrophoresis for inspection of DNA products

   

SECTION: For 2&#x27; PCR barcoded-head primers
11. Mix the 2' PCR master mixture gently by pippeting. Quick spin the tube.
SECTION: For 1&#x27; PCR head-primers
3. Mix the 1' PCR master mixture gently by pippeting. Quick spin the tube.
SECTION: For 1&#x27; PCR head-primers
6. Mix the reaction mixture gently by tapping the tubes. Quick spin the tubes.
SECTION: For 1&#x27; PCR head-primers
9. Markdown wells and upload the pictures to the Lab Google drive

  

SECTION: For 2&#x27; PCR barcoded-head primers
13. Add 2.5 µL pre-mixed barcoded-head primers (Forward + Reverse) to each PCR tube

SECTION: For 2&#x27; PCR barcoded-head primers
14. Add 1 µL of 1' PCR product as template, resulting in 25 µL reaction mixture for 2' PCR.

SECTION: For 2&#x27; PCR barcoded-head primers
15. Mix gently by tapping the tubes. Quick spin the tubes.
SECTION: For 2&#x27; PCR barcoded-head primers
17. Carry out electrophoresis for inspection of DNA products
SECTION: For 2&#x27; PCR barcoded-head primers
18. Markdown wells and upload the pictures to the Lab Google drive
1. Wear glove, clean up the working bench w. 1% bleach
SECTION: For 1&#x27; PCR head-primers
5. Add 0.6 µLDNA template in 8-strip PCR tubes, resulting15.6 µL reaction mixture for 1' PCR.
TITLE: KAPP-Sen TMC: Term Placenta Subject Selection Criteria
AUTHORS: Sonja Suvakov, Elizabeth Ann L. Enninga, Bridget A. Schley, Janelle Santos, Vesna D. Garovic

[DESCRIPTION]
Patient samples will be obtained during an elected cesarean (c-section) delivery. We will target two groups: Young (n=20, ages 20-30 years) and Mature (n=20, ages 40-50 years). We will enroll an equal number of male and female fetuses.

[STEPS]
1. Inclusion
Between the age of 20-30 or 40-50 years of age
Currently having a single birth (e.g. one baby)
Pregnancy is the result of natural conception
Elected for physician to perform a cesarean section for delivery
Pregnancy reaches full term (≥ 37 weeks)
2. Exclusion
Pregravid BMI > 35
Any health conditions including diabetes mellitus, cardiovascular disease, hypertension, or inflammatory disease
Any history of preeclamptic or complicated pregnancy prior to current pregnancy
TITLE: Artificial saliva
AUTHORS: Bjørn Peare  Bartholdy, a.g.henry

[DESCRIPTION]
Creating an artificial saliva solution for oral biofilm growth, specifically dental calculus.

This is a modified version of Sissons et al. 1991.

[BEFORE_START]
Mix the solution under a fumehood. It can smell pretty bad.
Also have the incubator under a fumehood, if possible. I had no such luxury...

[GUIDELINES]
This protocol explains how to mix 1000 mL  of artificial saliva.

[STEPS]
1. Add 300 mL distilled (or deionized) dH2O to a 1000 mL  beaker, with stirring and heat 60 °C .

2. Add:

-  2.5 g   
-  5 g   
- 10 g   
- 5 g   

Let the reagents completely dissolve before continuing to the next step

   

3. Add:

- 2.5 g   
- 0.35 g   
- 0.2 g   
- 0.74 g   
- 0.54 g   
- 2.5 mg   

4. Add the remaining 700 mL  distilled (or deionized) H2O

   

5. Adjust to pH 7 with   and stirring

6. Transfer to two 1000 ml bottles, so around half of each bottle is filled. 

Autoclave at 121 °C ,   for 15 min minutes
 

7. Once the solution has cooled, combine solutions to a single 1000ml bottle, and add:

1 mg   
0.3 g   
0.17 g   

 



8. Store in fridge at ca. 4 °C 

Occasionally test the pH of the solution to ensure it stays around pH 7
TITLE:  SYBR green RT-PCR assay  for the surveillance of SARS-CoV-2 variants of concern
AUTHORS: Pedro Cardoso, Fernando do Couto Motta, Marilda Agudo Mendonça Teixeira de Siqueira, Daniela Tupy de Godoy, Rodrigo Brindeiro, Monica Barcellos Arruda, Elisabete Andrade, Marisa Ribeiro, Marcela Fontana-Maurell, Elaine Costa, Daniele Rocha, Patricia Alvarez

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">The emergence of SARS-CoV-2 variants with multiple shared mutations has been described to be more transmissible and could affect COVID-19 morbidity and mortality. Some of these variants, known as B.1.1.7 (originally described in the United Kingdom), B1.351 (originally described in South Africa), and P.1 (originally described in Brazil) have rapidly become dominant within their countries and require a vigorous public health response. Whole Genomic sequencing remains the gold standard method to identify the SARS-CoV-2 variant, even though this approach is laborious, time-consuming, and expensive. Here, we have developed a fast and simple SYBR green-based real-time RT-PCR assay that identifies a distinct signature affecting the Non-Structural Protein 6 (NSP6), a nine-nucleotides deletion leading to amino acid losses: Δ106S, Δ107G, and Δ108F. This NSP6 signature is present in the variants of concern (VOC) described above (B.1.1.7; B1.351; P.1) and has not been detected in other SARS-CoV-2 lineages. The new version has corrections made on the master mix and authorship. </div></div>

[STEPS]
?. Briefly vortex and centrifuge reagents before use.
?. Prepare 10 µM working stocks of the primers.
?. Thaw the GoTaq® qPCR Master Mix (Promega), Nuclease-Free Water, and primers working stocks.
?. Use the 10 µM working stocks to prepare the mix, containing:Volume (per reaction)final concentrationGoTaq® qPCR Master Mix, 2X7.5 µl1xGoScript™ RT Mix for 1-Step RT-qPCR (50X)0.3 µl1xPrimer Forward0.15 µl0.1 µMPrimer Reverse0.15 µl0.1 µMNuclease-free water1.9 µl
Volume (per reaction)final concentrationGoTaq® qPCR Master Mix, 2X7.5 µl1xGoScript™ RT Mix for 1-Step RT-qPCR (50X)0.3 µl1xPrimer Forward0.15 µl0.1 µMPrimer Reverse0.15 µl0.1 µMNuclease-free water1.9 µl
?. Add 10µl to each weel or PCR tube.
?. Add 5 µl of RNA from positive controls 1 and positive control 2 to the correspondent well or PCR tube. Add 5 µl nuclease-free water to the negative control well or PCR tube. Mix by pipetting.
?. Add 5µl of extracted RNA (unknown sample) to the designated wells and mix by pipetting.
?. Seal plates and tubes.
?. Run the PCR with the following cycler conditions
?. Data Evaluation
Results Interpretation:Size of the fragmentTmNSP6: ΔS106; ΔG107 e ΔF108 60 bpNSP6: Wild type69bp≥74°C
TITLE: Novel Clinical Prediction Model: Integrating A2DS2score with 24-hour ASPECTS and Red Cell Distribution Width for EnhancedPrediction of Stroke-Associated Pneumonia following Intravenous Thrombolysis
AUTHORS: Sarawut Krongsut, Atiwat Soontornpun, Niyada Anusasnee

[DESCRIPTION]
Background:
Stroke-associated pneumonia (SAP) is a common leading cause of death during the acute phase. The A2DS2 score has been widely used to predict the risk of SAP. However, 24-hour non-contrast computed tomography-Alberta Stroke Program Early CT Score (NCCT-ASPECTS) and red cell distribution width (RDW) were not included in this scale. The purpose of the present study was to investigate the prognostic added value of combining 24-hour NCCT-ASPECTS and RDW with the A2DS2 score.
Methods:
A retrospective study of thrombolyzed acute ischemic stroke (AIS) patients from January 2015 to July 2022. Data on A2DS2 scores, 24-hour NCCT-ASPECTS, and RDW were collected. Three logistic regression models were created: Model A used only the traditional A2DS2 score; Model B (A2DS2-c) calculated probabilities using a logistic equation; and Model C (combined A2DS2-MFP) used multivariable fractional polynomial logistic regression and incorporated the A2DS2 score, 24-hour NCCT-ASPECTS, and RDW. Ischemic brain lesions in the middle cerebral artery area were assessed using 24-hour NCCT-ASPECTS after completing 24-hour intravenous thrombolysis.
Results:
Among a cohort of 345 thrombolyzed AIS patients, 70 individuals (20.3%) experienced SAP. The area under the receiver operating characteristic (AuROC) of 24-hour NCCT-ASPECTS and RDW were 0.841 and 0.621, respectively. The combined A2DS2-MFP calculation was significantly superior to the traditional A2DS2 score and A2DS2-c calculation (AuROC 0.917 vs. 0.880, P=0.026, and 0.917 vs. 0.888, P=0.024).
Conclusion: This study found that the 24-hour NCCT-ASPECTS and RDW enhanced the predictive value of the A2DS2 score for SAP after IV-tPA. The combined A2DS2-MFP model performed excellently in predictive performance, offering robust early SAP detection and potentially improving patient survival. Implementing this novel model in resource-constrained clinical settings could aid clinicians in effective monitoring, enabling risk stratification to guide clinical management.

[STEPS]
1. Novel Clinical Prediction Model: Integrating A2DS2
score with 24-hour ASPECTS and Red Cell Distribution Width for Enhanced
Prediction of Stroke-Associated Pneumonia following Intravenous Thrombolysis
TITLE: Sleep-Wake Recording in Dan lab
AUTHORS: Xiaolin Huang

[DESCRIPTION]
Surgery and Setups for Recording 24-Hour Natural Sleep/Wake Cycles in Mice.

[STEPS]
SECTION: Surgery
1. Mice were anesthetized with 1.5% isoflurane and placed in a stereotaxic frame. 

SECTION: Surgery
2. Body temperature was maintained using a heating pad. 

SECTION: Surgery
3. The skin was incised to expose the skull after asepsis and connective tissue was
removed.

SECTION: Surgery
4. To implant electroencephalogram (EEG) and electromyogram (EMG) recording
electrodes, two stainless steel screws were inserted into the skull 2.5 mm from
the midline and 3 mm posterior to the bregma, and two EMG electrodes were also
inserted into the neck muscles. 

SECTION: Surgery
5. A reference screw for grounding was placed on top of the left cerebellum. 

SECTION: Surgery
6. Insulated leads from the EEG and EMG electrodes were soldered to a pin header, which was
secured to the skull using dental cement.

SECTION: Recording
7. After a minimum of 7 days of undisturbed post-surgery recovery, behavioral
experiments were carried out in home cages placed in sound-attenuating boxes. 

SECTION: Recording
8. EEG and EMG electrodes were connected to flexible recording cables via a
mini-connector. 

SECTION: Recording
9. Recordings started after 1-2 days of habituation and continued for 3-4 days on a 12-h
dark/12-h light cycle. 

SECTION: Recording
10. Mouse behavior was recorded using a video camera at 10 frames per second (fps), along
with EEG and EMG recordings. 

SECTION: Recording
11. The signals were recorded with a TDT RZ10x/PZ5-32 or LR10-SK1 system.
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
A collection of protocols designed to guide the user in processing a viral metagenome from raw sequence data to assembly, and subsequent analysis. The user uses <em>actual</em> reads from <a href="https://github.com/MicroB3-IS/osd-analysis" target="_blank">Ocean Sampling Day (2014)</a> and processes them entirely within Cyverse, a NSF-supported cyberinfrastructure.

[STEPS]
?.
TITLE: Mouse Perfusion
AUTHORS: Haley   Geertsma

[DESCRIPTION]
This protocol is used to perfuse mice and isolate their brain for cryosectioning.

[STEPS]
1. Sedate mice with 120mg/kg Euthanyl then open their chest cavity to expose their heart.
2. Insert the needle tip into the left ventricle and cut the right atria. Begin slowly injecting 10mL 1X phosphate buffered saline (PBS) over 5 minutes.
3. Once all of the 1X PBS has been perfused, begin injecting 10mL 4% paraformaldehyde or 10% formalin over 5 minutes.
4. Extract brain and incubate them in either 4% paraformaldehyde or 10% formalin for 72 hours at 4oC.
5. Incubate brain in 10% sucrose for 24 hours at 4oC.
6. Incubate brain in 20% sucrose for 24 hours at 4oC.
7. Incubate brain in 30% sucrose for 24 hours at 4oC.
8. Decant brain from sucrose solution and flash freeze in isopentane at -35 to -45oC for 1 minute.
9. Cryo-section brain at 40μm.
TITLE: Gibson Protocol
AUTHORS: N.J. Hillson

[STEPS]
?. Prepare 6 ml of 5X ISO Buffer in a 15 ml falcon tube as follows:3 ml 1 M Tris-HCl pH 7.5+ 150 ul 2 M MgCl2+ 240 ul 100 mM dNTP mix (25 mM each: dGTP, dCTP, dATP, dTTP)+ 300 ul 1 M DTT+ 1.5 g PEG-8000+ 300 ul 100 mM NAD+ _______dH2O to6 mlStore at -20 C in 320 ml aliquots.
?. Prepare 1.2 ml of Gibson assembly master mix as follows:320 ul 5X ISO Buffer+ 0.64 ul 10 u/ml T5 exonuclease*+ 20 ul 2 u/ml Phusion polymerase+ 160 ul 40 u/ml Taq ligase+ _______dH2O to1.2 mlStore at -20 C in 15 ml aliquots.*This is optimized for 20-150 bp sequence homology overlaps
?. Thaw a 15 ml aliquot of the Gibson assembly master mix and keep on ice until use.
?. Measure the DNA concentration (ng/ml) of each assembly piece.
?. Add 100 ng of the linearized vector backbone and equimolar amounts of the other assembly pieces to the thawed 15 ml master mix in a 20 ml total volume assembly reaction mixture as follows: linearized vector backbone (100 ng)+ each additional assembly piece (to equimolar with backbone)+ 15 ml Gibson assembly master mix+ _______dH2O to20 ml
?. Incubate the assembly reaction at 50 C for 60 minutes, and then place on ice.
?. Transform 5 ml of the assembly reaction into 100 ml of competent E. coli and/or run a diagnostic agarose gel to check for successful assembly.
TITLE: Collecting Duckweed Samples For Microbial Community Profiling
AUTHORS: Kenneth Acosta

[STEPS]
?. [Processing Water Samples]
Get 150 mL 0.2 micron filter unit. Lay sterilized miracloth on top of filter unit.
?. [Processing Water Samples]
Collect 150-300 mL of water sample and pour through sterilized Miracloth into filter unit.
?. [Processing Water Samples]
Filter water sample through 0.2 micron filter unit by turning vaccum on.
?. [Processing Water Samples]
Cut 0.2 micron filter from unit using sterile scalpel.
?. [Processing Water Samples]
Place into 5 mL centrifuge tube.
?. [Processing Water Samples]
Flash freeze and store at -80C until processing.
?. [Processing Rinse Solution And Duckweed Tissue Samples]
Collect 1-3 grams of fresh weight Duckweed from environment sample. (See “Separating Duckweed From Other Solids” to obtain pure Duckweed tissue from environment.)
?. [Processing Rinse Solution And Duckweed Tissue Samples]
Place into 50 mL falcon tube.
?. [Processing Rinse Solution And Duckweed Tissue Samples]
Wash tissue 2x’s with sterile water.
?. [Processing Rinse Solution And Duckweed Tissue Samples]
Rinse tissue 3-5x’s with 30-40 mL rinse solution. Filter rinse solution through 0.2 micron filter each time.
?. [Processing Rinse Solution And Duckweed Tissue Samples]
Filter a total of 100-150 mL of rinse solution through 0.2 micron filter.
?. [Processing Rinse Solution And Duckweed Tissue Samples]
Cut filter using sterile scalpel, transfer to 5 mL centrifuge tube, flash freeze, and store for 16S rRNA amplicon sequencing.
?. [Processing Rinse Solution And Duckweed Tissue Samples]
Wash rinsed Duckweed tissue 2x’s with sterile water.
?. [Processing Rinse Solution And Duckweed Tissue Samples]
Transfer tissue to 5 mL centrifuge tube.
?. [Processing Rinse Solution And Duckweed Tissue Samples]
Flash freeze and store for 16S rRNA amplicon sequencing.