eamag/protocols-with-steps huggingface dataset:
TITLE: DNA extraction from plants
AUTHORS: Remco Stam

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Widely used protocol to extract DNA from plant leaves.</div><div class = "text-block">Many versions circulate on the web, this is the version as we use it.</div><div class = "text-block">It works well on maize, tomato and probably many other plants.</div></div>

[STEPS]
?. In a 2 ml tube, add 800 uL of 1,5x CTAB and 1 ul of Beta-mercaptoethanol to the ground leaf materialIncubate 1 hour at 60-65 degrees (C)
?. Cool at Room TempAdd 1 volume of Chloroform/Isoamylalkohol (24:1) mixtureMix on overhead shaker for 10 minutesCentrifuge at 3000 rmp for 25 minutes
?. Transfer supernatant to a new tubeUse wide pipette tips (cut point of tips if needed)
?. Add 10 uL RNAse (10 mg/ml)incubate at 37 degrees (C) for 30 minutes
?. Add one volume 100% Isoponanol (pre-chilled at -20)Allow sample to precepitate for up to 20 minutes at -20
?. Centrifuge: full speed, 10 minutesRemove supernatant
?. Add 1 ml 70% EthanolCentrifuge: full speed, 10 minutesRemove SupernatantRepeat once
?. Dry pellet in air or approx 20 minutes in vacuum centrifuge
?. Dissolve pallet in 1x TE (10 mM Tris, pH8, 1 mM EDTA)
TITLE: Joint angle calculation from optical markers and IMUs attached to the lower body
AUTHORS: Wolfgang Teufl, Markus Miezal, Bertram Taetz, Michael Fröhlich, Gabriele Bleser

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">The attached file contains a detailed description of the methods used in the manuscript "Validity of inertial sensor based 3D joint kinematics of static and dynamic sport and physiotherapy specific movements" for the calculation of 3D joint angles incorporating inertial sensor data and optical motion capture data.</div></div>

[STEPS]
TITLE: KAPP-Sen TMC: Tissue blocks paraffin embedding by JAX Histology Services (RNase-free)
AUTHORS: Anne Marchini, Tim Adams, Elaine Bechtel, Lesley Bechtold

[DESCRIPTION]
This protocol describes embedding of formalin-fixed tissues in paraffin (FFPE). Paraffin-embedded blocks are then kept at 4 °C and protected from light until RNA quality and spatial omics assays are performed.

[STEPS]
1. Wear prescribed PPE throughout all of the procedures described in this document. Change gloves if they get wet.
2. Histology does not process tissue by hand for RNase free requests. The usual tissue processing unit (Leica Histocore Peloris 3) is used. Use RNase free best practices when handling the tissue (wear gloves that have been wiped with RNAse Zap wipes) to load the tissue onto the processor and then into the embedding station.
3. Place cassettes containing tissue samples into a clean and empty paraffin bath within the embedding unit (Sakura Tissue-Tek TEC5 Embedding Console System). To avoid contamination, do not immerse in liquid paraffin within the embedding center.
4. Wipe gloves, molds and forceps with RNase Zap wipes before use. Allow the molds and forceps to dry completely. Use a pair of clean forceps for each tissue type. Do not re-use molds without cleaning between samples.
5. After the blocks are cooled and removed from the embedding molds, don gloves cleaned with RNase Zap wipes.  Then place the blocks in a plastic bag labeled with the job number and store them in a 4 °C refrigerator protected from light until they are sectioned.
6. Store paraffin blocks in a 4 °C refrigerator prior to cutting; RNA continues to degrade.
TITLE: DNA-Extraction-ClostridiaProject
AUTHORS: Tobias Wandt

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Protocol used for DNA-Extraction of Clostridia in my project. </div><div class = "text-block">DNA-Kit used is GenFind V3 by Beckmann/Coulter.</div><div class = "text-block">Some steps used from:</div><div class = "text-block">Sim JH, Anikst V, Lohith A, Pourmand N, Banaei N. Optimized Protocol for  Simple Extraction of High-Quality Genomic DNA from Clostridium  difficile for Whole-Genome Sequencing. J Clin Microbiol. 2015;53(7):2329-2331. doi:10.1128/JCM.00956-15</div></div>

[STEPS]
?. [Growing and Preparing Clostridia]
Inoculate Clostridia in 10ml TSB-Broth, incubating at least 48h in anaerobe atmosphere at 37°C
[TSB broth]
[at 37°C anaerobic]
?. [Growing and Preparing Clostridia]
Take 2ml of Clostridia TSB-Broth and centrifuge for 15min at 3,000g, after that remove supernatant
[Clostridia TSB broth]
[centrifuge at 3000g]
?. [DNA Extraction]
Add 500ul Lysis-Buffer(LBB) and 30ul Proteinase K, tipmixing or vortexing to resuspend the pellet
[LBB]
[Proteinase K]
?. [DNA Extraction]
Incubate at 57°C for 1h while continously shaking at ~500/min
[at 57°C + shaking 500/min]
?. [DNA Extraction]
Add 300ul vortexed Black Beads(BBB) and tipmix(at least 10 times), incubate for 5 min at room temperature, place on a magnetic tube stand until no or few beads are left in suspension ~10-15min, then carefully remove supernatant without disturbing the beads-formation
[Black Beads]
[at room temperature]
[magnetic tube stand]
?. [DNA Extraction]
Add 800ul WBB and tipmix(at least 10 times), place on a magnetic tube stand until no or few beads are left in suspension ~5min, remove supernatant
[WBB]
[magnetic tube stand]
?. [DNA Extraction]
[for a total of 2 WBB washes]
?. [DNA Extraction]
Add 800ul WBC and tipmix (at least 10 times), than add 600ul WBC and tipmix, place on a magnetic tube stand until no or few beads left in suspension ~5min , remove supernatant
[WBC]
[magnetic tube stand]
?. [DNA Extraction]
[for a total of 2 WBC washes]
?. [DNA Extraction]
Add 100ul distilled water, tipmix 10 times, incubate for 2min at roomtemp, tipmix 10 times again and go to next step OR place in a 4°C fridge overnight, tipmix 10 times next day
[distilled water]
[room temperature]
[at 4°C]
?. [DNA Extraction]
Place on a magnetic tube stand until no beads are left in suspension
?. [DNA Extraction]
Take the supernatant with your DNA and put it in a fresh tube
TITLE: Long Primer PCR (for Trypanosoma brucei) 
AUTHORS: Alex  Zegarra

[STEPS]
?. PRC Mix  1 uL pPOT (25 ng/uL)0.2 mM dNTPs1 uM for primer 1 uM rev primer 1 uL PCR grade DMSO 5uL 10x buffer 2 (Roche) XX uL ddH20 for total volume of 49 uLAdd 1 uL Expand High Fidelity polymerase (Roche) once mixture has reached 94 C.
?. Maintain procyclic form SMOX P9 cells [31] between 1x106 -1x107  cells ml-1 for at least 72 hours prior to transfection to ensure they are in log growth phase.
?. PCR conditions 94 C  5 mins 94 C 15 sec 65 C 30 sec   (30 cycles) 72 C 2 min 72 C 7 min
?. Centrifuge 1x107 log phase procyclic cell per transfection at 800 g for 10 min at room temp
?. remove all supernatant
?. Resuspend cells in 500 mL of room temperature cytomix per transfection, and add to a 4 mm gap electroporation cuvette
?. Add 50 mL of unpurified PCR to the cell suspension
?. Electroporate the cell once with 1.7 kV, 25 mF (gene pulser (Bio-Rad) or three times 1.7 kV for 100 Ms, 200 ms interval (BTX ECM830 (hardvard Apparatus))
?. Recover the cells for 8-16 hours in 10 ml SDM-79 at 28 C
?. Add the appropriate selective drug to the final concentrations:Blasticidin(Melford)20mg/ml(Hygromycin b Gold (Invivogen)25mg/ml(>25mg.ml-1reduces transfection efficiency due to low readthrough transcription of the resistance cassette).
?. If clones are required, dilute 5 ml of recovered cells into 50 ml of selective medium and distribute 1 ml aliquots into a 48 well plate.
?. Resistant populations of cells emerge after 7 – 10 days, and clones emerge after 10 – 14 days.
?. FOLLOW UPTransfection of bloodstream formT. bruceiusing theAmaxaNucleofector IIT. bruceican be efficiently transfected using theAmaxaNucleofector II using the human T-cell kit (VPA-1002 Lonza).
?. 1. Completethe human T-cell Nucleofector solution by addition of supplement 1. The combined solution and supplement can be stored and is stable for 3 months at 4°C.
?. Purify 100ml of long primer PCR (~8mg) with onephenol chloroform extractionfollowed by ethanol precipitation at -80°C for 1 hour with two 70% ethanol washes (the pellet should be easily visible). Note that using a silica membrane column instead of phenol chloroform to purify the DNA will reduce transfection efficiency by 10 –50 fold.
?. Resuspend the dried pellet in 10ml 5mM Tris pH8.
?. Maintain bloodstream form SMOX B4 cells[31]between 1x105–1x106cells.ml-1for at least 72 hours prior to transfection to ensure they are in log growth phase.
?. Centrifuge 2×107log phase (
?. Carefully remove all supernatant.
?. Resuspend the cell pellet in 100ml of completeAmaxaT cell buffer per transfection, and transfer to anAmaxacuvette.
?. Add the purified DNA to the cell suspension, and electroporate once using Program X-001.
?. Recover cells for 8 - 16 hours in 50 ml HMI-9 at 37°C with 5% CO2.
?. Add the appropriate selective drug to the final concentrations:Blasticidin(Melford)5mg/ml. Hygromycin B Gold (Invivogen)1.5mg/ml.
?. Distribute 1ml aliquots of cells into two 48 well plates.
?. Resistant clones emerge after 6 – 8 days.
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
This is the protocol for dephosphorylation of 5'-ends of DNA using AnP (Antarctic Phosphatase - M0289).

[GUIDELINES]
<p><strong>Dephosphorylation of 5' -ends of DNA in Restriction Enzyme Reaction</strong></p>
<ul>
<li>The phosphate can be added directly into the digestion reaction during or after DNA digestion</li>
<li>Antarctic Phosphatase is active in all NEB restriction enzyme buffers only when supplemented with Antarctic Phosphatase Reaction Buffer, which provides Zn2+ required for enzyme activity</li>
<li>The restriction enzyme should be heat inactivated at the same time as the phosphatase after digest and dephosphorylation</li>
<li>If restriction enzyme cannot be heat inactivated, DNA purification is required before ligation</li>
</ul>

[STEPS]
?. 
?. 
?.
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p>Cara memasukkan foto mentah (<em>raw</em>) ke dalam <em>photo frame</em> yang sudah disiapkan dalam format PowerPoint, agar siap dipublikasi di media sosial dan situs web.</p>

[STEPS]
?. 
?. 
?. 
?. 
?. 
?.
TITLE: Total and PS129 aSyn levels using Western Blot
AUTHORS: daniel.dautan daniel, Wojciech Paslawski, Per Svenningsson

[DESCRIPTION]
Measure total and ps129 alpha-synuclein in mouse plasma. 

[STEPS]
1. Dilute plasma samples 10-fold in MilliQ water. 
2. Mix the sample in a 1:1 ratio with 20 mM TCEP to reduce disulfide bridges, and then 1:1 with a denaturing loading buffer (50 mM Tris-HCl, 70 mM Tris, 1% lithium dodecyl sulphate (LDS), 5%
glycerol, 0.25mM EDTA, 0.11mM SERVA Blue G-250, 0.0875mM Phenol Red, pH-8.5). Boil at 95°C for 5 min.
3. Separate 20 μL of each sample using a 4-12% bis-tris acrylamide gel using MES running buffer (50mM Tris, 50mM 2-(N-morpholino)ethanesulfonic acid (MES), 0.1% SDS, 1mM EDTA, pH-7.3). 
4. After Sodium dodecyl sulphate polyacrylamide gel electrophoresis, incubate gels in a transfer buffer (25mM Tris, 192mM glycine, 30% methanol) and 0.45μm pore size Immobilon-P‱ PVDF membranes
(Millipore, MA, US). Pre-wet membranes in methanol.
5. Assemble gels with membranes. Perform transfer using the Trans-Blot Turbo Transfer System (BioRad, CA, US) according to manufacturer protocols.
6. Dry membranes and fix quickly in 100% methanol.
7. Between each incubation period, membranes were
washed three times in Tris Buffered Saline (TBS, 20mM Tris, 150mM NaCl, pH-7.6)
containing 0.1%(v/v) Tween20 (TBS-T).
8. Block membranes for 60 min in 5% skim milk at Room temperature . 
10. Incubate membranes for 120 min with appropriate HRP-conjugated secondary antibody (Dako/Agilent, CA, US) diluted 1:10000(v/v) in 1% skim milk at RT.

11. Develop the signal with Clarity‱ Western ECL Substrate (Bio-Rad, CA, US). Visualize levels of  proteins by measuring band intensities with ImageJ.
12. Normalized target protein amounts to total protein in the sample, measured by BCA assay according to manufacturer instructions (Thermo Fisher Scientific, MA, US). 
13. Use fluorescent scan to visualize the protein ladder.
9. Incubate membranes 60 min at 4 °C with a primary antibody diluted 1:1000(v/v) in 1% skim milk. 

Antibodies used were:
-Anti-Total-Alpha-Synuclein Polyclonal Antibody (Thermo Fisher Scientific, PA5-143581, MA, US) 
-Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (Abcam, ab51253, UK)
TITLE: Culturing Chlamydomonas reinhardtii
AUTHORS: Joao Vitor Molino

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><span>This protocols describe the steps required for the growth of </span><span style = "font-style:italic;">Chlamydomonas reinhardtii</span><span> in liquid media.  </span></div></div>

[STEPS]
?. [Material]
TAP media or other (How to prepare TAP media here)Erlenmeyer flaskOrbital shakerLight source
?. [Inocullum]
*Inoculation can be perfomed by scraping an agar plate containing Chlamydomonas reinhardtii  or from another liquid cultureInnoculate the cells in an erlenmeyer containing media (Ex:   containing   of TAP media or  containing )Place the flask in a shaker at   with  and with   of incident white light ( to   works)Time to stationary phase depends on initial innoculum.
[Erlenmeyer flask]
50 mL
[Erlenmeyer flask]
250 mL
25 33
[of orbit]
80
60
120
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
This protocol outlines our analysis of the potential antibiotic resistance genes found within the skin virome. We start by visualizing the relative abundances of the top 10 antibiotic resistance categories (according to the CARD). We then quantify the number of unique antibiotic resistance gene categories annotated in the samples at each skin site, and also look at the percent CARD annotated ORFs of the overall ORFs present in each sample at each skin site.

[BEFORE_START]
Supplemental information available at:<br /><br /><a href="https://figshare.com/articles/The_Human_Skin_dsDNA_Virome_Topographical_and_Temporal_Diversity_Genetic_Enrichment_and_Dynamic_Associations_with_the_Host_Microbiome/1281248" target="_blank">https://figshare.com/articles/The_Human_Skin_dsDNA_Virome_Topographical_and_Temporal_Diversity_Genetic_Enrichment_and_Dynamic_Associations_with_the_Host_Microbiome/1281248</a>

[GUIDELINES]
<span class="identifier">sessionInfo</span><span class="paren">(</span><span class="paren">)<br /><br />## R version 3.2.0 (2015-04-16) <br />## Platform: x86_64-apple-darwin13.4.0 (64-bit) <br />## Running under: OS X 10.10.4 (Yosemite) <br />## ## locale: <br />## [1] en_US.UTF-8/en_US.UTF-8/en_US.UTF-8/C/en_US.UTF-8/en_US.UTF-8 <br />## <br />## attached base packages: <br />## [1] stats graphics grDevices utils datasets methods base <br />## <br />## loaded via a namespace (and not attached): <br />## [1] magrittr_1.5    formatR_1.2    tools_3.2.0    htmltools_0.2.6 <br />## [5] yaml_2.1.13    stringi_0.4-1    rmarkdown_0.7    knitr_1.10.5 <br />## [9] stringr_1.0.0    digest_0.6.8    evaluate_0.7</span>

[STEPS]
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?.
TITLE: ELISA for anti-SpA antibodies in hyper-immune egg whites.
AUTHORS: Angel  Justiz-Vaillant

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

[STEPS]
?. The 96 well polystyrene microplate (U-shaped bottom) is coated with 500 ng of SpA (Sigma-Aldrich) in coating buffer for 4 h at 37oC.
?. The microplate is washed four times with PBS-Tween-20 and blocked with 3% non-fat milk in PBS, 25 μl/well, 1h, RT.
?. The microplate is washed four times again.
?. Samples of egg whites are added in 1:10 dilutions.
?. After incubation for 1h at RT the microplate is washed four times.
?. Then, 50 μl of peroxidase-labelled protein-A diluted 1:3000 (Sigma-Aldrich) is added.
?. The microplate is incubated for 1h at RT.
?. The microplate is washed four times.
?. Tetramethylbenzidine (TMB) solution (50 μl) is added to each well.
?. Perform a further incubation of 15 min in the dark
?. The reaction is stopped with 3M H2SO4.
?. Read the microplate  in a microplate reader at 450 nm.
?. Calculate the cut-off point as appropriate.
TITLE: Kujawinski Lab cleaning protocol
AUTHORS: Krista Longnecker

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">This protocol describes lab cleaning procedure for glass and plastics.</div></div>

[STEPS]
?. [Kujawinski Lab cleaning protocol]
Add a small amount of diluted Citranox to hot water in tub (less than into ~2 gallons of water).
5 mL
?. [Kujawinski Lab cleaning protocol]
Add dishes to the tub making sure every surface is covered with water.
Containers with chunky bits should be scrubbed with a brush to remove all evidence of particulate material.
?. [Kujawinski Lab cleaning protocol]
Let sit .
?. [Kujawinski Lab cleaning protocol]
Rinse thoroughly with hot tap water.
?. [Kujawinski Lab cleaning protocol]
After rinsing, place containers into acid bath and let sit .
?. [Kujawinski Lab cleaning protocol]
Rinse thoroughly with MilliQ water.
?. [Kujawinski Lab cleaning protocol]
Allow to dry.
?. [Kujawinski Lab cleaning protocol]
Cover open tops and exposed surfaces with aluminum foil.
?. [Kujawinski Lab cleaning protocol]
For combusting (glassware only):
?. [Kujawinski Lab cleaning protocol]
Combust for at least  at .
450 °C
?. [Kujawinski Lab cleaning protocol]
For autoclaving:
?. [Kujawinski Lab cleaning protocol]
Autoclave for .
TITLE: Quality Control and Quality Assurance (Part 11 of Safety and Efficacy of Imatinib for Preserving Beta-Cell Function in New-onset Type 1 Diabetes Mellitus)
AUTHORS: Stephen.Gitelman , Jeffrey A. Bluestone

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">This is Part 11 of "Safety and Efficacy of Imatinib for Preserving Beta-Cell Function in New-Onset Type 1 Diabetes Mellitus".</div><div class = "text-block">This clinical study is supported by JDRF. The aim of the collection is to determine whether imatinib will slow the progression of the autoimmune destruction of ß cells and lead to the preservation of C-peptide secretion in T1DM and to assess Diabetes-related objectives and safety of Imatinib in new-onset type 1 diabetes mellitus".</div></div>

[STEPS]
TITLE: Magic Red Cathepsin B activity assay
AUTHORS: Razaul Karim

[DESCRIPTION]
This protocol details the magic red Cathepsin B activity assay.

[STEPS]
SECTION: Cell Culture/Treatments
1. 0.5x10^4/ml of cells plated on coverslips.

SECTION: Cell Culture/Treatments
2. PFF-488 treatment.

SECTION: Cell Culture/Treatments
3. PBS/trypsin (0.01%) wash.

SECTION: Cell Culture/Treatments
4. 1x Magic Red solution for last 15 min at 37 °C in cell culture incubator.

SECTION: Imaging
5. Live-Cell imaging.

SECTION: Imaging
6. Leica Stellaries 8 confocal laser scanning microscope, assembled with live cell chamber with CO2 gas, temperature and humidity controlled.

SECTION: Imaging
7. Image analysis:
Image J. quantification of the % area of the cell covered by Magic Red signal, as well as total signal intensity per cell.
TITLE: 6M GITC
AUTHORS: Jacquelina.Woods  

[DESCRIPTION]
This method was developed at the FDA’s Center for Food Safety and Applied Nutrition for GenomeTrakr’s pandemic response project, monitoring SARS-CoV-2 variants in wastewater​​. Protocols developed for this project cover wastewater collection, concentration, RNA extraction, RT-qPCR detection, library prep, genome sequencing, quality control checks, and data submission to NCBI. This method provides a reagent formula required in the extraction of RNA from viral concentrates using the RNeasy and Zymo kits. 

[STEPS]
1. Aseptically mix 7.09 g  in sterile distilled or ultrapure water to 4.5 mL

2. Bring total volume up to 10 mLwith additional sterile distilled or ultrapure water 

3. Store at 2-8 Room temperaturein the dark (or in light occluding tube)
TITLE: Protocol for preparing post-mortem tissue for intracellular neuromelanin quantification in H&amp;E-stained sections.
AUTHORS: Marta Gonzalez-Sepulveda, Thais Cuadros, Miquel Vila

[DESCRIPTION]
Protocol for preparing post-mortem tissue for intracellular neuromelanin quantification in H&E-stained sections

[STEPS]
SECTION: Tissue Processing
1. Formalin-fixed paraffin-embedded tissue blocks from the pontine and midbrain regions were sectioned at 5µm, collected onto adhesive microscope slides and allowed to dry at room temperature for 24 hours. 
SECTION: Tissue Processing
2. Slides were incubated in the oven at 60°C for 30 minutes to melt the paraffin.
SECTION: Tissue Processing
3. To remove the paraffin, sections were submerged in Xylene (Panreac Applichem 211769.2714) for 3 x 3 minutes, followed by rehydration in decreasing ethanol concentrations (100% ethanol for 2 x 5 minutes, 95% ethanol for 2 x 5, 70% ethanol for 2x5 minutes) and distilled H2O for 5 minutes. 
SECTION: Tissue Processing
4. Standard hematoxylin-eosin (H&E) staining.
SECTION: Image Acquisition
5. Sections were scanned using 20x objective (NA=0.8) with pre-set focusing and exposure parameters for optimal neuromelanin signal quality with an automated Slide Scanner Olympus (SLIDEVIEW VS200, Tokyo, Japan).
SECTION: Image Acquisition
6. Identical parameters were applied to each scanned section to ensure consistency in capturing neuromelanin.
TITLE: Pre-extraction sample processing for CALIBER project
AUTHORS: James JN Kitson

[DESCRIPTION]
This is a document that outlines the steps needed to prepare field-collected samples during the CALIBER project for archiving and further processing.

[STEPS]
SECTION: Setup
1. Source steel beads (ball bearings) for tissue grinding (Tungsten beads are not usually necessary). We use hardened carbon steel or stainless steel bearings from simplybearings.co.uk. This protocol requires two beads per sample tube.
SECTION: Setup
2. Beads are usually shipped coated in manufacturing oil (especially the carbon steel beads). To remove this, place beads in a borosilicate glass beaker or Duran bottle with the pouring lip and lid removed then bake for at least 12 hours at 250 oC.

  

SECTION: Setup
3. Prepare 5 ml screwcap collection tubes (e.g. Starlab #E1450-1100) containing three 3 mm hardened steel beads in batches of 96 tubes.

  

SECTION: Insect sample processing
4. Pick out a frozen plant sample bag from the freezer and note the sample name.


  

SECTION: Archive plant sample processing
8. Write the sample name on a manilla sample drying envelope and find the corresponding dried sample label.

  

SECTION: Insect sample processing
5. Using clean forceps, pick off all insects and place in a screwcap tube of the appropriate size (2ml/5ml/15ml) then fill the tube with a solution of 95% Ethanol and 5% Glycerol.
SECTION: Insect sample processing
7. Store at -20 oC until transported to SASA for qPCR analysis.

  

SECTION: Insect sample processing
6. Label the tube with the corresponding tube label.


    

SECTION: Archive plant sample processing
9. Into this envelope, place approximately 2-5 g of leaf and chopped stem material.
SECTION: Archive plant sample processing
10. Seal with the dried sample label and place in an oven overnight at 35-40 oC with a tray of silica gel in the bottom.

  

SECTION: Archive plant sample processing
11. The archive sample is ready for long term storage at room temperature.
SECTION: DNA extraction plant sample processing
12. Using a clean scalpel, slice and place ~ 50 mg of plant material into the pre-prepared collection tubes containing the hardened steel ball bearings (from step 3). 
SECTION: DNA extraction plant sample processing
13. Label the tube with the corresponding tube label.

  

SECTION: DNA extraction plant sample processing
14. Store at -20 oC until ready to proceed with DNA extraction.
SECTION: Cleanup and reset
15. The remaining plant material and packaging can be disposed of in a clinical waste bin.
SECTION: Cleanup and reset
16. Wash forceps and scalpel in 1x chemgene.
SECTION: Cleanup and reset
17. Rinse forceps and scalpel in molecular grade water.
SECTION: Cleanup and reset
18. Rinse forceps in 100% Isopropanol and leave to dry.
SECTION: Cleanup and reset
19. Wipe down dissection area with blue roll and 1x chemgene.
SECTION: Cleanup and reset
20. Proceed with the next sample.