eamag/protocols-with-steps huggingface dataset:
TITLE: An analysis of Relative Telomere Length (RTL) during peri-operative chemotherapy in patients with operable gastric or gastro-oesophageal junction adenocarcinoma
AUTHORS: Jeff Evans, Nicol Keith

[DESCRIPTION]
OBJECTIVES
The primary objective of this study is:
(a) to analyse Relative Telomere Length (RTL) in blood samples taken from patients during peri-operative chemotherapy for operable gastric or gastro-oesophageal junction adenocarcinoma. This will allow us to determine if the baseline (pre-treatment) RTL correlates with patient outcome as measured by relapse-free (RFS) and overall survival (OS).
The secondary objectives of this study are:
(a) to determine if the baseline (pre-treatment) RTL correlates with “tumour response” as measured by down-staging of the primary tumour with the pre-operative component of chemotherapy (comparison of the radiological staging at diagnosis, before pre-operative chemotherapy, with pathological staging at subsequent resection or with repeat radiological staging in the event of progressive disease so that the tumour is inoperable).
(b) to analyse changes in markers associated with cellular senescence in blood samples taken from patients during peri-operative chemotherapy for operable gastric or gastro-oesophageal junction adenocarcinoma. These will include cathelicidin-related antimicrobial protein (CRAMP), EF-1a, stathmin, and chitinase 3-like protein 3.
The tertiary (exploratory) objectives of this study are:
(a) to analyse changes in CK-18 in blood samples taken from patients during peri-operative chemotherapy for operable gastric or gastro-oesophageal junction adenocarcinoma as a marker of drug-induced cell death by apoptosis.
(b) to analyse changes in senescence-associated micro-RNAs in blood samples taken from patients during peri-operative chemotherapy for operable gastric or gastro-oesophageal junction adenocarcinoma.

STUDY DESIGN
This will be a multi-centre, open, non-randomised study. Eligible patients will be those with operable gastric, gastro-oesophageal junction or lower oesophageal adenocarcinoma who are about to undergo peri-operative chemotherapy with either the ECF/EOF or ECX/EOX (epirubicin, Cisplatin/oxaliplatin and capecitabine/5-FU) regimens. Additionally, patients randomised in the NCRN STO3 study to receive ECX + bevacizumab will also be eligible.

[STEPS]
TITLE: Step A: Culturing
AUTHORS: k.z.cooper

[DESCRIPTION]
Culturing step for bacterial genome resequencing project

[STEPS]
SECTION: Preparation
2. Ensure agar plates are pre-poured
SECTION: Preparation
3. Pre-label the plates using only the numbers provided on the spreadsheet under heading ‘Sample_Label’
SECTION: Preparation
4. Get required glycerol stocks and keep on dry ice
SECTION: Creating streak plate
6. Create streak plate inoculum using inoculation loop from glycerol stock
SECTION: Creating streak plate
5. Ensure to cross reference with spreadsheet before every inoculation
SECTION: Creating streak plate
7. Replace inoculation loop (after step I in the below figure) and dilute across the plate
SECTION: Creating streak plate
9. Incubate at 37°C overnight
SECTION: Creating LB broth overnight culture
10. Pre-label tubes using numbers from spreadsheets
SECTION: Creating LB broth overnight culture
11. Transfer 5 ml of LB broth into a universal tube
SECTION: Creating LB broth overnight culture
12. Inoculate the LB broth with a single colony from the agar plate
SECTION: Creating LB broth overnight culture
13. Grow at 37°C overnight at 180 rpm in a shaking incubator
SECTION: Creating LB broth overnight culture
14. Place agar plates into fridge for storage
SECTION: Creating streak plate
8.   

1. Open the tracking form and verify you also have a copy of the batch tracking sheet printed out:
Step A tracking form
Batch tracking sheet
TITLE: Single Nucleus RNAseq Sample Prep from Nodose Ganglia
AUTHORS: Sebastian  Preissl, Jamie   Verheyden, Xin Sun

[DESCRIPTION]
How to isolate single nucei from nodose vagal ganglia neurons in preparation for 10XGenomics sequencing.

[STEPS]
1. Nodose ganglia are dissected and flash frozen in liquid nitrogen.Store at -80C until ready to isolate nuclei.Grind up the sample with liquid nitrogen using the 1.5mL centrifuge tube and fitting pestle.

2. Add 500 ul Lysis Buffer to pulverized tissue, pipette 10x up and down and place on ice
Incubate 5 min with the overhead shaker in the cold room
3. Spin down 5 min 500g at 4C using soft settings. Remove supernatant
4. Resuspend tissue in 400 ul Sorting Buffer by pipetting 10x up and down.
5. Filter with a green 30 um CellTRic (Sysmex) into FACS tube. Wash filter with another 200 ul SB. Gently clap rack against bench to fully transfer the nuclei suspension. Add 6 ul DRAQ7 (Cell signaling) to stain nuclei and place sample on ice.
6. For collection of sorted nuclei add 50 ul Collection Buffer

 to a 1.5 ml LoBind tube (collection tube).
7. Use 100 um Chip for sorting of nuclei
Set temperature for sample and collection to 5C
Identify single nuclei based FSC/SSC signals andDRAQ7 fluorescence in FL-6 (PE-Cy7) channel
8. Sort 60,000 (200 ul) nuclei (Gate: “Sorting”) with the setting “Purity” into the prepared collection tube.  After sorting is finished (10-20 min), mix sample and keep on ice.
9. Spin down 15 min 1000xg at 4C (Using swinging bucket rotor). After centrifugation, there should be a tiny blue pellet be visible. Remove supernatant and resuspend in 18 ul RB by pipetting 10x (Recovery of nuclei should be around 50-60%). Leave sample on ice.
10. 

Check for single cell suspension and count nuclei using hemocytometer. Mix 5 ul sample + 5 ul Trypan blue and count 4 big squares. 
Calculation: (counted number)/4*20=nuclei/ul). Adjust concentration to 1000 nuclei/ul.  Follow 10xGenomics library prep protocol.
TITLE: HLA genotyping using SS-SBT methods
AUTHORS: Ryosuke Tashiro, Hidetoshi Inoko, Kuniyasu Niizuma, Teiji Tominaga

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><span>Genomic DNA was obtained from the patients’ 2ml of whole blood using the QIAamp DNA Mini Kit for genomic DNA purification (Qiagen GmbH, Hilden, Germany), and 400 ng of purified genomic DNA was used for polymerase chain reaction (PCR) amplification. DNA was preserved in 4°C freezer. The basic cycling parameters were as follows: (i) first denaturation at 94°C for 2 min, followed by 30 cycles of denaturation at 98°C for 10 s and 60°C for 20 s and extension at 68°C for 5 min (</span><span style = "font-style:italic;">HLA-A</span><span>, </span><span style = "font-style:italic;">HLA-B</span><span>, and </span><span style = "font-style:italic;">HLA-C</span><span>); (ii) first denaturation at 94°C for 2 min, followed by 30 cycles of denaturation at 98°C for 10 s and annealing at 70°C for 5 min (</span><span style = "font-style:italic;">HLA-DRB1</span><span>and </span><span style = "font-style:italic;">HLA-DPB1</span><span>); and (iii) first denaturation at 94°C for 2 min, followed by 30 cycles of denaturation at 98°C for 10 s and annealing at 70°C for 9 min (</span><span style = "font-style:italic;">HLA-DQB1</span><span>). Long-range PCR reactions were performed using the thermal cycler Gene Amp PCR System 9700 (Life Technologies, Carlsbad, CA, USA). The PCR products obtained were purified with Agencourt AMPure XP (Beckman Coutler, CA, USA) and quantified by the Quant-iT Picogreen dsDNA Assay Kit (Thermo Fisher Scientific, MA, USA). Next, the PCR products were clonally amplified and barcoded using the Ion Plus Fragment Library Kit (Life Technologies), and the barcoded library was sequenced using the Ion Torrent Personal Genome Machine DNA sequencing system (Life Technologies). The NGS read data were analyzed by Sequence Alignment Based Assigning Software (SeaBass), and finally, the </span><span style = "font-style:italic;">HLA</span><span>alleles were determined.</span></div></div>

[STEPS]
?. DNA extraction
?. Long-ranged PCR
?. Sequencing
?. Data analysis
?. Construction of barcoded library
?. Fisrts denature
94 °C
?. Denature
98 °C
?. Annealing (30 cycles)
[HLA-A, B,C]
[HLA-DRB1]
[HLA-DQB1]
[HLA-DQB1]
[HLA-A, B, C]
[HLA-DRB1]
?. Output of NGS read data
?. Homology search using Blat
?. Selection of allele candidates
?. Mappng of reads and candidate allele sequence
?. Calculation of coverage
?. Final confirmation
?. Preparation of the Enriched Template-Positive Ion Sphere particles (ISPs)
?. Extension (30cycles)
[HLA-A,B,C]
[HLA-A,B,C]
TITLE: Confocal imaging power settings – protocol and discussion
AUTHORS: Marco Polin, Antoine Allard, Kelsey Cremin, Emily Skates, Orkun S Soyer

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Confocal microscopy is a commonly used microscopy technique involving a point laser scanned across a sample for generating a pixel-by-pixel image of (auto)fluorescent samples (see here for a useful explanatory video and slide set on confocal microscopy). Two key parameters in confocal microscopy are the power of the laser used, and the ‘scan time’. The latter determines how long the laser ‘dwells’ at each point on the sample. Together, these two parameters determine how much photons the sample is exposed to. In turn, the amount of photon the sample receives contributes to phototoxicity and photobleaching effects. Phototoxicity refers to inhibitory/damaging effects of light on cell physiology, while photobleaching relates to (auto)fluorescent molecules losing their ability to emit photons. Both phototoxicity and photobleaching are crucial factors in image-based, live-cell analyses, as we do not want to perturb our samples unnecessarily and we want the highest possible signal. While zero phototoxicity and photobleaching are physically impossible, we aim to reduce these effects as much as possible. It is therefore crucial that we understand the laser settings when conducting confocal microscopy. This protocol will;</div><div class = "text-block">• motivate you to care about laser settings,</div><div class = "text-block">• introduce how to set them, and</div><div class = "text-block">• describe how to report them</div></div>

[STEPS]
?. [Confocal imaging power settings-step-by-step protocol]
Prepare sample.
?. [Confocal imaging power settings-step-by-step protocol]
Use sample to set your microscope up for correct laser and scanning settings.
?. [Confocal imaging power settings-step-by-step protocol]
Make sure you use a calibrated power-meter and that it is zeroed before measurement and set to the correct wavelength matching that of your laser.
?. [Confocal imaging power settings-step-by-step protocol]
Remove sample and place power meter on to the sample holder or place power-meter slide, in case you are using a power-meter embedded in a slide.
?. [Confocal imaging power settings-step-by-step protocol]
Turn laser on and record power measurement.
?. [Confocal imaging power settings-step-by-step protocol]
Repeat measurement few times to make sure you get an average / approx. value at andaround the specimen position.
?. [Confocal imaging power settings-step-by-step protocol]
BE AWARE of lasers, and do not look into light path, while holding the power-meter in place.
?. [Confocal imaging power settings-step-by-step protocol]
Record laser power value and perform experiment.
?. [Confocal imaging power settings-step-by-step protocol]
At end of experiment, repeat 3 to 8 again and record laser power for end of experiment.
TITLE: High Spatial Resolution MALDI Imaging Mass Spectrometry Data Acquisition
AUTHORS: Katerina V Djambazova, Martin Dufresne, Angela R.S.  Kruse, Jamie Allen, allison.b.esselman, Madeline E. Colley, David       Anderson, ali.zahraei, Olof  Isberg, Melissa Farrow, Jeff Spraggins

[DESCRIPTION]
This protocol provides the workflow and instrument parameters for setting up a MALDI imaging mass spectrometry experiment on a Bruker timsTOF Flex instrument. This protocol is intended for 10µm spatial resolution imaging of lipids (m/z 400 - 2000) in positive and negative ionization mode data acquisition in qTOF mode of operation.

[STEPS]
1. Place slides in a MTP 2-slide holder. Scan slides using flatbed scanner, allowing for sufficient contrast to visualize the tissue boundary. Ensure the slides have fiducials.
 

4. Use the FlexImaging software to load the optical image of the slides. Teach the target position with three teaching points. Check the accuracy of the teaching by moving the stage to a fourth location. 
3. Calibrate the TOF with an external calibration using red phosphorus. Proceed if all calibration points are <1ppm standard deviation error and the calibration score is >99%.
 

5. Select desired tissue measurement region. Start data acquisition in the FlexImaging software.
2. Load the 2-slide holder in the instrument. Open timsControl software and select appropriate MALDI IMS data acquisition method. Generate target profile by ensuring "Use Target Profile" is enabled. Check the height at a few different locations on the slide and ensure the difference is <10 µm.
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p style="text-align: center;"><img class="s-mce-img" style="display: block; margin-left: auto; margin-right: auto;" src="https://s3.amazonaws.com/pr-journal/hnnfjtn.png" width="100" height="90" data-src="https://s3.amazonaws.com/pr-journal/hnnfjtn.png" data-ofn="nehirarma-üst-Tr.png" />  </p>
<p style="text-align: center;">Nehir Biyoteknoloji Ltd. <a href="http://www.nehirbt.com.tr/en/" target="_blank">www.nehirbt.com</a></p>

[BEFORE_START]
<p>This protocol is derived from</p>
<p>[Reference: Processing guidelines for permanent epoxy negative photoresist SU8 2025, SU8 2035, SU8 2050 and SU8 2075, MicroChem company<strong>]</strong></p>

[STEPS]
?. 
?. 
?. 
?.
TITLE: Lithium Acetate / SDS Extraction of Genomic DNA from Saccharomyces cerevisiae
AUTHORS: Clark Fritsch

[DESCRIPTION]
This protocol is just a quick description of the protocol provided by Looke et al., 2011 (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3182553/). The purpose of this protocol is simply to provide a quick and easy method to isolate genomic DNA from yeast for PCR-based applications. While it does not provide the purist genomic DNA that you can get, it is extremely quick and easy to perform.

[STEPS]
1. Pick one yeast colony from the plate or spin down 100-200 μl of liquid yeast culture (OD600=0.4). Suspend cells in 100 μl of 200mM LiOAc, 1 % SDS solution. Alternatively, you can spin down 100 - 500 uL of liquid culture at 3,000 rcf for 4 minutes and then resuspend the pelleted cells in 100 uL of 200 mM LiOAc, 1% SDS solution if you wish.
2. Incubate for 5 minutes at 70°C.
3. Add 300μl of 96-100 % ethanol, vortex thoroughly.
4. Spin down DNA and cell debris at 15,000 rcf for 3 minutes.
5. Resuspend the pellet in 70% ethanol. Then spin the resuspended DNA and cell debris mixture at 15,000 rcf for 3 minutes.
6. Dispose of the supernatant and then dissolve the pellet in 100 μl of H2O or TE and spin down cell debris for 15 seconds at 15 000 g.
7. Use 1 μl of supernatant for PCR.
TITLE: Pulldowns 
AUTHORS: Dan Tudorica

[DESCRIPTION]
Traditional in vitro pulldown

[STEPS]
SECTION: Pulldown
1. In a final volume of 30 μL, 20 μg of purified MBP-Rubicon RH domain was mixed with 20 μg of RAB7A in 50 mM HEPES 7.5, 150 mM NaCl, 2 mM MgCl2, 10 mM TCEP buffer.

Include a negative control consisting of only soluble Rab7, with no MBP-Rubicon, in order to test your washing efficacy.
SECTION: Pulldown
2. Incubate samples on rocker at room temperature for 1 H
SECTION: Pulldown
3. Add 20 uL of 50% v/v amylose resin to each sample
SECTION: Pulldown
4. Allow to rock for an additional 30 min to bind
SECTION: Pulldown
5. Collect resin at bottom of tube via a tabletop centrifuge, aspirate off and discard supernatant, and wash with 500 uL of ice cold buffer for three total washes.
SECTION: Pulldown
6. Elute sample by resuspending beads in 20 uL of buffer + 20 mM maltose for 10 min on shaker.
SECTION: Pulldown
7. Prepare samples for SDS-PAGE by adding 1x loading buffer, and load onto a 4-12% gel. Run gel at 120 V until dye front reaches the bottom of the gel
SECTION: Pulldown
8. Stain with Coomassie blue G-250, and destain with water until bands are clearly visible. Image.
TITLE: Setting up a liquid culture and harvesting C.elegans
AUTHORS: Vidur Sabharwal

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

[STEPS]
?. Add 30ml S Medium to a sterilized 250 ml flask.
?. Inoculate the S Medium with a concentrated E. coli OP50 pellet to a final concentration of 30g/L
?. Wash each of 5 large plates (90cm) of C. elegans (just cleared of bacteria) with 5 ml S Medium and add to the 30 ml flask.
?. Put the flask on a shaker at 200rpm
20 °C
?. Put the flask on ice to allow the worms to settle.
?. Aspirate most of the liquid from the flask.
?. Transfer the remaining liquid to a 15 ml borosil conical tube using a glass pasteur pipette and spin for at least 2 min at 500 × g to pellet the worms. Young larvae may take longer than 2 min to pellet.
?. Aspirate the remaining liquid and wash with 10x volume of M9 twice. Spin for at least 2 min at 500 × g to pellet the worms
?. Transfer the worms to a 1.5ml eppendorf, incubate on ice for 10 mins and remove as much supernatant as possible.
?. Add IP Buffer to the Pellet with 2 parts homogenization buffer : 1 part worm pellet and store at -80C for upto 3 months
?. Transfer 20 L4 hermaphrodites each to 10 90cm plates. They will starve out in approximately 6 days. Start the following steps on the day the plates are almost starved.
20 °C
?. After the second wash step, animals were resuspended in 20 ml of ice-cold M9 lacking MgSO4, followed by the addition of 20 ml of ice-cold 60 % sucrose in H2O.
?. After vigorous mixing of the sucrose/worm mixture, 4 ml of ice-cold M9 lacking MgSO4 was gently layered on top, and the worms were centrifuged at 400 g for 2 min at 4 °C.
The sucrose float steps were performed as quickly as possible, as otherwise, the layer of animals failed to form properly.
A layer of animals was now visible on top of the sucrose, while contaminants sedimented at the bottom.
?. To maximize recovery, 30 ml of supernatant was aspirated from the sucrose float, and distributed into four 50 ml tubes that were subsequently filled by addition of room-temperature M9 lacking MgSO4. The room temperature M9 allowed the animals to digest OP50 bacteria in their intestine.
?. The four tubes were placed on ice to cool down for 30 min, after which the animals were washed twice in M9 buffer. Add IP Buffer to the Pellet with 2 parts homogenization buffer : 1 part worm pellet and store at -80C for upto 3 months
TITLE: Use of cholera toxin subunit B to label neural projections to lower urinary tract organs
AUTHORS: Janet R Keast, Peregrine B Osborne, John-Paul  Fuller-Jackson

[DESCRIPTION]
This protocol is used to visualise sensory and autonomic neurons innervating organs of the lower urinary tract in an experimental adult male or female rat. The protocol is performed under anesthesia and should incorporate all local requirements for standards of animal experimentation, including methods of anesthesia, surgical environment, and post-operative monitoring and care.

[STEPS]
SECTION: Preparation for surgery
1.  Prepare cholera toxin subunit B solutions: low salt formulation with 0.05% Evans Blue.
SECTION: Preparation for surgery
2. Anesthetise animal (2.5% isoflurane in oxygen, or as required for maintenance)
SECTION: Preparation for surgery
3.  Apply eye lubricant and place animal on heated pad.
SECTION: Preparation for surgery
4.  Shave and clean the ventral abdomen.
SECTION: Surgery
5.  Perform a midline incision in the skin and then the muscle, then gently move organs to visualise the required injection site.
SECTION: Surgery
6. Microinject sterile tracer solution at the selected injection site using a Hamilton Neuros Syringe attached to a 33G needle. At each injection site, hold the needle in place for ~5 seconds after ejection of the dye, to enable the dye to spread to the underlying tissue. This also minimises leakage.
 

SECTION: Surgery
7.  Wash all injection sites with sterile saline.
SECTION: Surgery
8.  Close the muscle and skin using approved procedures. Administer analgesics and monitor animal during postoperative period as per local approved procedure. 
SECTION: Tissue harvesting
9. To analyse tracer distribution in ganglia or the injection site, 4 days after surgery, fix animals by intra-cardiac perfusion, then remove tissues of interest for further study.
TITLE: Sequencing Protocols for the One Thousand Plant Transcriptomes Initiative
AUTHORS: Eric J. Carpenter, Naim Matasci, Shuangxiu Wu, Jing Sun, Jun Yu, Fabio Rocha Jimenez Vieira, Chris Bowler, Richard G. Dorrell, Matt Gitzendanner, Ling Li, Wensi Du, Kristian Ullrich, Michael S. Barker, James H. Leebens-Mack, Gane Ka-Shu Wong

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">The 1000 Plants (1KP) initiative explored the genetic diversity of green plants (Viridiplantae) by sequencing RNA from 1,342 samples representing 1,173 species. All of the analyses done for the 1KP capstone, and previous studies on subsets of these data, are based on a series of de novo transcriptome assemblies and related outputs that will be described in this publication. We expect that these data will also be useful to other researchers with interests in specific gene families, either across the green plant tree of life or in more focused lineages. These protocols provide additional details on the sample and sequence processing, hopefully providing insight for reproducibility and tools for other researchers working on plant transcriptomics and evolution.</div></div>

[STEPS]
TITLE: Food intake behavior protocol
AUTHORS: Santiago Unda, Michael G. Kaplitt

[DESCRIPTION]
This protocol describes the behavioral test to measure food intake in mice.

[STEPS]
1. Habituate mice in single cages 2 days prior the test day.
2. Fast the mice for 18 hours with access to water.
3. Using a small weight scale measure food chow and allow ad libitum food intake.
4. Record food chow weight at 1, 2-, 4-, 6-, and 24-hours post-food chow access.
TITLE: qPCR: Bacterial SSU rRNA 338F-516P-805R
AUTHORS: Roey Angel, Eva Petrova, Ana Lara

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Universal 16S rRNA probe-based-qPCR assay for bacteria.</div><div class = "text-block"><span>The primers and probe are taken from </span><a href="http://dx.doi.org/10.1002/bit.20347" style = "text-decoration:underline;color:blue;cursor:pointer;"><span style = ":;">Yu et al. (2005)</span></a><span style = ":;">.</span></div></div>

[STEPS]
?. [Primers and probe]
ABCD1NameTypeSequenceTarget region12BAC338FForwardACT CCT ACG GGA GGC AG338-3543BAC516P2ProbeTGC CAG CAG CCG CGG TAA TA516-5364BAC805RReverseGAC TAC CAG GGT ATC TAA TC 785-8051. Relative to E. coli SSU rRNA gene2. The probe must be dual-labelled either with 5’-6-FAM, 3’-BHQ1 or any other valid combination
ABCD1NameTypeSequenceTarget region12BAC338FForwardACT CCT ACG GGA GGC AG338-3543BAC516P2ProbeTGC CAG CAG CCG CGG TAA TA516-5364BAC805RReverseGAC TAC CAG GGT ATC TAA TC 785-805
?. [qPCR mixture]
ABCD1ReagentFinal concentration1 tube (20 μl)plate (20 μl x 100)2PCR H2O&nbsp;2.246032x TaqMan Fast Advanced Master mix1x1010004BSA (20 μg μl-1)0.4 μg μl-10.4405338F0.5 μM1.01006805R0.5 μM1.01007516P0.2 μM0.4408Template&nbsp;55 x 100
ABCD1ReagentFinal concentration1 tube (20 μl)plate (20 μl x 100)2PCR H2O&nbsp;2.246032x TaqMan Fast Advanced Master mix1x1010004BSA (20 μg μl-1)0.4 μg μl-10.4405338F0.5 μM1.01006805R0.5 μM1.01007516P0.2 μM0.4408Template&nbsp;55 x 100
?. [Thermocycler programme]
for         2.1  for         2.2  for       }
95 °C
95 °C
62 °C
[take snapshot]
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
Tris-Acetate-Phosphate medium (TAP) is among the most common for Chlamydomonas reinhardtii. This protocol uses premade TAP from <a href="https://www.thermofisher.com/order/catalog/product/A1379801" target="_blank">ThermoFisher</a>. Alternative and significantly cheaper TAP medium can be found at <a href="http://phytotechlab.com/index.php/tris-acetate-phosphate-tap.html" target="_blank">PhytoTechnology</a>. Agar is from Acros.

[STEPS]
?. 
?. 
?. 
?.