eamag/protocols-with-steps huggingface dataset:
TITLE: Diagnosis and treatment of plasmablastic lymphoma, plasmablastic myeloma and plasmablastic neoplasm: protocol for a scoping review
AUTHORS: Hirotaka Mori, Masahiko Fukatsu, Hiroshi Ohkawara, Yuka Oka, Yuki Kataoka, Shunsuke Taito, Takayuki Ikezoe

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><div class = "justify" style = "text-align:left"><span style = "font-weight:bold;">Introduction</span></div></div><div class = "text-block"><div class = "justify" style = "text-align:left">The differential diagnosis of plasmablastic lymphoma (PBL) form plasmablastic myeloma (PBM) is sometime very difficult. In such cases, the diagnosis of plasmablastic neoplasm (PBN) is acceptable. But clinicians face the question whether chemotherapies for PBL, PBM or others are effective for PBN and the question increases the diagnosis dilemma. Although evidence on the differential diagnosis and treatment of those three entities is compiling, there is no scopingreview of different diagnosis and treatment for PBL, PBM, and PBN to answer those questions. The aim of this study is to develop an a priori protocol to undertake a scoping review of the available evidence on the reproducibility of differential diagnosis and the treatment of PBL, PBM, and PBN. </div></div><div class = "text-block"><div class = "justify" style = "text-align:left"><span style = "font-weight:bold;">Methods and analysis</span></div></div><div class = "text-block"><div class = "justify" style = "text-align:left">For the conducting the scoping review protocol, we will use an scoping review methodology established in the Joanna Briggs Institute manual. This methodology has a five-stage approach: (1) identify the research question; (2) identify relevant studies; (3) select studies; (4) chart the data and (5) collating, summarize and report the results. We also will use the Preferred Reporting Items for Systematic Reviews and Meta-Analyses Extension for Scoping Reviews to assess the results.</div></div><div class = "text-block"><div class = "justify" style = "text-align:left"><span style = "font-weight:bold;">Ethics and dissemination</span></div></div><div class = "text-block">Since this is a review of the literature, ethics approval is not indicated. We will disseminate the findings from this study in publications in peer-reviewed journals.</div></div>

[STEPS]
TITLE: Sorbitol washing complex homogenate for improved DNA extractions
AUTHORS: Ashley Jones, Benjamin Schwessinger

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">The extraction of pure DNA can be challenging due to the presence of sugars, oils and other endogenous chemicals present within biological samples. This is particularly true for many plants and fungi due to the presence of secondary metabolites such as polyphenols and polysaccharides. Polyphenols within the cytosol can become irreversibly DNA-bound after cell lysis and polysaccharides can co-precipitate with DNA during the extraction. Sorbitol is an osmotically active sugar alcohol and washing homogenate with sorbitol before cell lysis has been shown to significantly improve the purity of DNA extractions. Sorbitol does not pass cell membranes and likely acts by drawing the cytosol out of the cell. Therefore polyphenols and polyscacharides would be removed.</div></div>

[STEPS]
?. [PREPARATION]
Prepare sorbitol wash solution. Approximately 2x volume of tube capacity will be used per sample. Sample tissue should not exceed 33% of tube capacity.To prepare a 500 mL solution:  ABCDE1ReagentTarget concentrationMolecular weightStock concentrationAdd from stock2D-Sorbitol0.35 M182.17powder31.88 g3PVP 40,0001% (w/v)40,000powder5 g4Tris-HCl pH 8100 mM157.601 M50 mL5EDTA pH 85 mM292.240.5 M5 mL6MQWNANANABring to 500 mL
ABCDE1ReagentTarget concentrationMolecular weightStock concentrationAdd from stock2D-Sorbitol0.35 M182.17powder31.88 g3PVP 40,0001% (w/v)40,000powder5 g4Tris-HCl pH 8100 mM157.601 M50 mL5EDTA pH 85 mM292.240.5 M5 mL6MQWNANANABring to 500 mL
Store at 4°C for up to 6 months.DTT or β-Mercaptoethanol will be added freshly when used (below).
?. [HOMOGENISATION]
Grind tissue to a fine powder, keep frozen with liquid nitrogen.
?. [SORBITOL WASH]
Fill the tube capacity with an excess of sorbitol wash solution.
?. [SORBITOL WASH]
Add DTT, approximate final concentration of 1 mM. Add 0.15 mg DTT per 1 mL of sorbitol wash. A tiny scoop of powder is sufficient. DTT MW = 154.253.
Alternatively add 1% β-Mercaptoethanol (v/v). Add 10 µL of β-Mercaptoethanol per 1 mL of sorbitol wash.
?. [SORBITOL WASH]
Shake, invert and vortex to mix thoroughly.
Ensure tissue is in suspension.
?. [SORBITOL WASH]
Centrifuge at 5,000 rcf for 5 min at room temperature.
Limited based on tubes rupturing if they contain ball bearings. Can also be reduced to 2,500 rcf.
?. [SORBITOL WASH]
Carefully decant the supernatant. Remove as much of the wash solution as possible without losing the pellet.
Supernatant should appear slightly cloudy or discoloured light yellow or brown.
?. [SORBITOL WASH]
If the supernatant was turbid, viscous or had dark discolouration, repeat the sorbitol wash.
?. [HOMOGENISATION]
If homogenate is not in a tube (i.e. mortar and pestle), transfer to an appropriate sized tube.
Homogenate should not exceed 33% of tube capacity.
?. [DNA EXTRACTION]
Proceed to a DNA extraction method of choice by adding lysis buffer to the pellet. Continue protocol as normal.
TITLE: Catalase
AUTHORS: Eva Feldman, Frank Brosius

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Summary: </span></div><div class = "text-block">Describes the protocol used by the DiaComp to detect and quantify catalase activity in a tissue. </div><div class = "text-block"><span style = "font-weight:bold;">Diabetic Complications:</span></div><div style = "text-align :; float : ;"><img style = "" src = "https://s3.amazonaws.com/pr-journal/bibp4kcn.jpg" /></div><div style = "text-align :; float : ;"><img style = "" src = "https://s3.amazonaws.com/pr-journal/bibq4kcn.jpg" /></div><div style = "text-align :; float : ;"><img style = "" src = "https://s3.amazonaws.com/pr-journal/bibr4kcn.jpg" /></div><div style = "text-align :; float : ;"><img style = "" src = "https://s3.amazonaws.com/pr-journal/bibt4kcn.jpg" /></div><div style = "text-align :; float : ;"><img style = "" src = "https://s3.amazonaws.com/pr-journal/bibu4kcn.jpg" /></div><div style = "text-align :; float : ;"><img style = "" src = "https://s3.amazonaws.com/pr-journal/bibv4kcn.jpg" /></div><div style = "text-align :; float : ;"><img style = "" src = "https://s3.amazonaws.com/pr-journal/bibw4kcn.jpg" /></div></div>

[STEPS]
?. Sample Preparation:
?. [TISSUE:]
1. Homogenate tissue in 1X Reaction Buffer on ice. 2. Using a black plate, pipette 25µL of diluted standards, controls (if any) and samples into wells. (Final concentration will be fourfold lower, 0 to 10µM) 3. Prepare stock solution of 20mM H²O² then prepare a 40µM H²O² dilution by adding 10µl of 20mM H²O² to 4.99mL 1X Reaction Buffer. 4. Pipet 25µL of 40µM H²O² solution into each well. 5. Incubate for 30 minutes at room temp. 6. Prepare stock solution of 10mM Amplex Red reagent and divide into 50µL aliquots and freeze immediately. 7. Prepare stock solution of 100U/ml HRP and divide into 20mL aliquots. 8. Prepare 100µM Amplex Red reagent containing 0.4U/mL HRP by adding 50µL of 10mM Amplex Red stock solution and 20µL of 100U/ml HRP stock solution to 4.93mL 1X Reaction Buffer. 9. Begin 2nd phase of reaction by adding 50µL of the above to each well. 10. Place plate into Fluroskan holder and click "START" . 11. Take 4 readings @ 15 minute intervals using 544/590 filter pairs. (Generally take 3 reading which would be after 30 min. incubation as recommended.) 12. Save raw data as an Excel file into the CTx data folder. Use the naming convention CTXXXX.xls, where XXXX is the date in mmdd format. 13. Select Process>Organize. Choose the appropriate data to organize (usually Measure1), then click "OK". This re-arranges the data into columns. 14. Save organized data as an Excel file into the Catalase data folder. Use the naming convention ctXXXXor.xls, where XXXX is the date in mmdd format.
TITLE: Detection of Cryptosporidium in stool samples by Taq-Man qPCR
AUTHORS: botchiesenyo  

[DESCRIPTION]
Detection of Cryptosporidium parvum and/or Cryptosporidium hominis from fecal samples by qPCR.

[STEPS]
SECTION: qPCR
3. Combine the following for a single qPCR sample (9μl  per sample). Scale accordingly for number of samples and technical replicates:

5 μl           10 x Luna Universal Probe qPCR Master Mix
0.25 μl      10 μM Forward Primer (ATGACGGGTAACGGGGAAT)
0.25 μl      10 μM Reverse Primer (CCAATTACAAAACCAAAAAGTCC)
1μl             1 μM Probe ([FAM]CGCGCCTGCTGCCTTCCTTAGATG[BHQ1])
2.5μl          Ultrapure water

SECTION: qPCR
4. Load mastermix in 96-well PCR plate using a multichannel pipette.

Add 1μl extracted DNA to corresponding wells.

Keep a detailed plate map of which wells contain standard curve DNA, sample DNA, positive control DNA, no DNA control.
SECTION: qPCR
5. Spin plate in a centrifuge at low speed to make sure the samples are at the bottom of the well.

SECTION: qPCR
6. Run qPCR cycling as described for Luna Universal qPCR Mixture. Make sure qPCR machine is measuring FAM at the correct cycle:





SECTION: qPCR
6.1. 95°C - 3 minutes
SECTION: qPCR
6.2. 
Repeat 40x:
95°C - 10 seconds
60°C - 30 seconds (record FAM in this step)

SECTION: DNA Extraction
2. Follow protocol from Quick-DNA Fecal/Soil Microbe Miniprep Kit from Zymo Research to extract total genomic DNA from stool sample (both standard curve samples and experimental samples).

Stool samples (0.1 g) should be thoroughly vortexed in lysis buffer and then subjected to five freeze-thaw cycles prior to DNA extraction.
SECTION: Generating samples for a standard curve
1. To 100 mg of fecal material (known to not be infected with Cryptosporidium), spike in a known number of Cryptosporidium parvum. 
SECTION: Calculating number of oocysts/g
7. Calculate delta Ct values for samples that contain the standard curve. 

Using Microsoft excel (or a similar graphing program) plot the delta Ct values on the Y-axis and the number of oocysts/g on the x-axis.

Use the analysis tools to determine the linear equation of your standard curve samples.
SECTION: Calculating number of oocysts/g
8. Use the equation determined from your standard control samples, you can estimate the number of oocysts/g in your experimental samples.

Y = delta Ct value for experimental sample
Solve for X = oocysts/g for your experimental sample
TITLE: Gibson Assembly
AUTHORS: Zhujun Wei

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">This protocol includes two different kits. Choose one to use.</div></div>

[STEPS]
?. Set up a small box with ice, put DNA and Gibson Mix into it before going into the metal bath or the Bio-rad S1000TM thermocycler cycler .
?. Add the following reagent to a PCR tube (4 μl).  AB1Gibson Mix2 μl2insertX μl3vectorY μl X+Y=2 ul , X : Y=3 : 1~5 : 1.Temperature : Time= 50℃ : 15-30 min
AB1Gibson Mix2 μl2insertX μl3vectorY μl
?. Set up a small box with ice, put DNA and Gibson enzyme and Gibson buffer into it before going into the metal bath or the Bio-rad S1000TM thermocycler cycler .
?. Add the following reagent to a PCR tube (10 μl).  AB1Gibson enzyme1 μl2Gibson buffer2μl3insertX μl4vectorY ulX=0.02× vector length /vector concentrationY=0.02× insert length /insert concentrationTemperature : Time= 37℃ : 30-60 min
AB1Gibson enzyme1 μl2Gibson buffer2μl3insertX μl4vectorY ul
TITLE: Aptamer conjugated beads affinity assay
AUTHORS: Jorge Fernández

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">The following protocol details how to test affinity by an aptamer-conjugated particles with their targets, via target immobilization in 96-treated wells.</div></div>

[STEPS]
?. [Well Coatings]
Add to each well of the 96 maxisorp plate 100 µL of E. Coli or Streptavidin-DNA suspensions. And incubate overnight at 4 ºC.
?. [Well Coatings]
After incubation, remove the liquid in the plate flipping the 96-plate.q! Don't aspire the liquid in the wells with a pipette, to avoid damaging the coating.
?. [Well Coatings]
Add 150 µL of 3% BSA solution to each well. Make two adittional BSA adittions for negative controls. Incubate the 96-wells for 4h with mild agitation.
?. [Affinity Assay]
Add 80 µL of each conjugated latex beads stock to the coated wells. Remember to add at least in two wells, coated with the different targets respectively; E. Coli and Streptavidin-DNA conjugate. Incubate under mild agitation at room temperature for 1 hour.
?. [Affinity Assay]
Remove the liquid by plate flipping, as mentioned in step 2.
?. [Affinity Assay]
Wash the wells adding 200µL PBS-T (Tween-20 0.05 %), incubating 2 minutes under mild agitation, and flipping the 96-plate. Repeat this step twice.
?. [Affinity Assay]
Cover the wells with 50 µL of distilled water and measure absorbance at the absorbtion peak for latex beads.For the the fluorescent orange latex beads we have employed, the absorbsnce measurement have been performed at 570nm.
TITLE: Testing of Total Dissolved Solids in Coffee Samples
AUTHORS: Shannon.logan

[DESCRIPTION]
Testing the total dissolved solids levels in different coffee samples

[STEPS]
1. Grind coffee to desired consistency and weigh 20g to put in cafetiere.
2. Boil kettle with 250ml of deionised water.
3. Pour boiling water into beaker and allow to cool to 93°C, using a digital thermometer to ensure
accuracy of temperature.
4. Once the water has cooled to the desired temperature, pour into the cafetiere and allow to brew for
4 minutes, using a timer/stopclock.
5. Once the coffee has brewed, use the cafetiere as stated in the instruction manual, and pour coffee
into beaker. 

6. Using a syringe, Take 1ml of the coffee and place in the refractometer, placing the syringe against the walls of the well to cool the sample for better reading. 
7. Take note of the refractometer reading.
8. Repeat this process for all coffee samples.
TITLE: Microfluidics 3 - Mold Fabrication: 3D Printing
AUTHORS: Serhat Sevli,  C. Yunus Sahan

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><div class = "justify" style = "text-align:left">Su8 is a photoresist resin used in MEMS technology for thick structures. PDMS microfluidic chips fabrication is suitable for SU8+Si wafer molds. This protocol describes the Direct UV laser lithography application performed in our laboratories.</div></div></div>

[STEPS]
?. [3D Printer Setup]
Molds for PDMS microfluidic chips can be fabricated using 3D Printing technology. SLA(/DLP) printers are mostly prefered for microfluidics due to its high resolution, relative low cost, and being suitable for direct microchannel formation.NehirBT uses a SLA/DLP 3D printing technology for transferring drawings by CAD software to channels directly as mold. The important parameters arranged are; SpecsInfoTotal size50x110x150Light sourceUV LED/DLPPoint size30-50 micronsVelocity 30 sec per layerResolution30-50 micronsAspect ratio1:1003D structuresunlimited
SpecsInfoTotal size50x110x150Light sourceUV LED/DLPPoint size30-50 micronsVelocity 30 sec per layerResolution30-50 micronsAspect ratio1:1003D structuresunlimited
?. [Microfluidic Channel Design]
The designs are generated using a CAD software in STL format and with milimeter as a unit.The flow simulation is performed using a multiphysics finite element analysis software, so the design optimization is performed by simulation support and correcting the errors. MiiCraft software is used for converting the design file to MII format which is suitable for MIICRAFT100 instrument.MiiCraft software controls the MIICRAFT100 instrument by arranging the duration of exposure per layer and layer thickness.
?. [3D Printing]
The resin choice is critical for the 3D Printing of PDMS molds since chemicals inside resin materials inhibit the curation of PDMS polymer. Only there are a few suitable resins and NehirBT uses; 1- OOkuma; FP60 functional prototyping resin, 2- ResinWorks; Master Mold Resin for PDMSResin is put inside the reservoir of the 3D printing device. The printing is initiated by the setup of SpecsInfoLayer thickness30-50 micronsLayer curing time1-3 secGap adjustment0,1 mmBase layer curing time1-20 secBase layer layer number1-10
SpecsInfoLayer thickness30-50 micronsLayer curing time1-3 secGap adjustment0,1 mmBase layer curing time1-20 secBase layer layer number1-10
?. [Post Fabrication Recipe]
After fabrication of the PDMS mold, it must be thermally and optically cured. Optical curing is exposure to UV LED (385-405 nm) for several hours and thermal treatment at 95oC for several hours is needed.
Not enough post-fabrication treatment causes remaining inhibitory chemicals which causes non-cured PDMS.
?. [Washing of 3D Printed Mold]
The prepared mold is washed with isopropanol after completing the treatments and then dried with pressurized nitrogen gas.The mold is now ready for PDMS microchannel manufacturing. Put the mold in a petri dish, and label it with all appropriate information. Pouring PDMS material on mold in a petri dish is performed in class2 laminar flow hood.
TITLE:       ELISA for quantification of IL-37 in human serum.
AUTHORS: Angel  Justiz-Vaillant

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Interleukins (IL) are a type of cytokine first thought to be expressed by leukocytes alone but have later been found to be produced by many other body cells. They play essential roles in the activation and differentiation of immune cells, as well as proliferation, maturation, migration, and adhesion. They also have pro-inflammatory and anti-inflammatory properties. The primary function of interleukins is, therefore, to modulate growth, differentiation, and activation during inflammatory and immune responses. Interleukins consist of a large group of proteins that can elicit many reactions in cells and tissues by binding to high-affinity receptors in cell surfaces. </div><div class = "text-block"><span>IL-37 plays an essential role in the regulation of the innate immunity causing immunosuppression. Phagocytes and organs including the uterus, testis, and thymus express it. IL-37 upregulates immune responses and inflammation in autoimmune disorders. </span><a href="https://www.ncbi.nlm.nih.gov/books/NBK499840" style = "text-decoration:underline;color:blue;cursor:pointer;"><span style = ":;">[1]</span></a></div><div class = "text-block">Reference</div><div class = "text-block"><span>1. Justiz Vaillant AA, Qurie A. Interleukin. In:</span><span style = "font-style:italic;">StatPearls</span><span>. Treasure Island (FL): StatPearls Publishing; June 12, 2019.</span></div></div>

[STEPS]
?. An anti-human IL-37 coating antibody is adsorbed onto the microwells by incubation overnight at 4°C with carbonate-bicarbonate buffer.
?. Add 50 µl of human serum. Human IL-37 present in the serum sample binds to antibodies adsorbed into the microwells.
?. The microplate is blocked with 3% non-fat milk-PBS buffer and later wash to remove unbound proteins.
?. Fifty (50) µl of biotin-conjugated anti-human IL-37 antibody is added. The optimal dilution must be investigated.
?. The microplate is rewashed with PBS-Tween 20 buffer, pH 7.4.
?. One hundred µl of streptavidin-HRP conjugate is added and it binds to the biotin-conjugated anti-human IL-37 antibody. The optimal dilution of this conjugate must be investigated.
?. The plate is washed following incubation to remove the unbound Streptavidin-HRP.
?. Add 100 µl of 3',3',5',5'- tetramethylbenzidine (TMB; Sigma-Aldrich) into each well.
?. Incubate the microwells in the dark for 15 min.
?. A colored product is formed in proportion to the quantity of human IL-37 present in the sample or standard.
?. The reaction is terminated by addition of 100 µl 3M H2SO4  and the absorbance is measured at 450 nm.
?. A standard curve is made from 7 human IL-37 standard dilutions and the human IL-37 sample concentration is determined.
?. For better results place the microplate on a microplate shaker in every incubation.
TITLE: Production of lentiviruses in Lenti-X cells
AUTHORS: Cole S Sitron, F Ulrich Hartl

[DESCRIPTION]
This is a protocol to produce lentivirus in a HEK cell line selected to produce high-titer virus.


[STEPS]
SECTION: Transfection
1. Plate Lenti-X cells 2.5 x 10^6/10 cm 1440 min before transfection. 

SECTION: Transfection
2. Warm OptiMEM, X-tremeGENE HP DNA Transfection Reagent, and DNA to warm to Room temperature.

SECTION: Transfection
3. Add 1 mL OptiMEM to a tube.

SECTION: Transfection
4. Add plasmid DNA to the OptiMEM, gently pipetting to mix.

 

SECTION: Transfection
5. Add X-tremeGENE HP DNA Transfection Reagent to OptiMEM/DNA (4 µL per 1 µg of DNA).

SECTION: Transfection
6. Incubate for 15 min at Room temperature.

SECTION: Transfection
7. Add OptiMEM/DNA/X-tremeGENE to the plate of cells dropwise, then gently swirl the plate.
SECTION: Transfection
8. The day after transfection, replace the medium with 
SECTION: Collection
9. 4320 min after transfection, collect the medium in a 15 ml conical.

SECTION: Collection
10. Centrifuge 500 x g x 5 min, then pass the supernatant through a   filter.

SECTION: Collection
11. Add 3 mL of LentiX concentrator to the supernatant, invert 6 times to mix, then incubate 45 min at 4 °C.

SECTION: Collection
12. Centrifuge at 1500 x g for 45 min at 4 °C.

SECTION: Collection
13. Remove the supernatant and resuspend the pellet in 120 µL ice cold PBS. 

SECTION: Collection
14. Make aliquots in multiples of 14 µL and freeze at -70 °C.
TITLE: diDO-IPTL protocol
AUTHORS: Jacob   Waldbauer, Lichun Zhang

[DESCRIPTION]
Isobaric peptide termini labeling (IPTL) is a quantification method which permits relative quantification using quantification points distributed throughout the whole tandem mass spectrometry (MS/MS) spectrum. It is based on the complementary derivatization of peptide termini with different isotopes resulting in isobaric peptides.

[STEPS]
SECTION: DAY 1
1. Prepare dried MS-grade trypsin, 2ug per sample to be labeled. Note that separate aliquots of dried trypsin must be used for 16O- and 18O-labeling.
SECTION: DAY 1
2. Prepare two separate N-methylmorpholine (NMM) and acetic acid O-isotope exchange buffers with 16O and 18O (>98%) enriched water. Volume ratio of buffer components is 48.5 H2O : 1 NMM : 0.5 acetic acid (pH 7.4). The total volume of buffer to prepare depends on how many peptide samples are to be labeled. The protocol below is for 40µL of O-isotope exchange buffer per sample (~10-20 µg of peptide). For samples with small amounts of peptide, the concentration can be raised by adding only 20µL buffer per sample and halving the volumes of all subsequent reagent additions.
SECTION: DAY 1
3. Redissolve dried trypsin aliquots in their respective O-isotope exchange buffers and transfer 40µl to each tube of dried peptide sample. KEEP TRACK of which tubes received which buffer.
SECTION: DAY 1
4. Parafilm the tops of the tubes and incubate at 37ºC overnight.
SECTION: DAY 2
5. After overnight incubation, add 2µl of 11.3M monochloroacetic acid (prepared in LC-MS grade water) to each peptide tube to lower pH down to 2.6.
SECTION: DAY 2
6. Prepare 16% CH2O and CD2O formaldehyde solutions, at least 2µl per sample, diluting stocks with LC-MS grade water if necessary.
SECTION: DAY 2
7. Prepare 4.8M NaBH3CN in LC-MS water. Tare 1.5mL tube, add a small amount of NaBH3CN powder in fume hood, weigh, dissolve.
SECTION: DAY 2
8. Add 2µL of CH2O to each 18O tube and 2µL of CD2O to each 16O tube.
SECTION: DAY 2
9. Add 2µL 4.8M NaBH3CN to each tube.
SECTION: DAY 2
10. Mix well and incubate at 45ºC for 1 hr. While waiting, prepare 5M ammonium formate in LC-MS water.
SECTION: DAY 2
11. Add 2µL 5M ammonium formate to each tube and mix well.
SECTION: DAY 2
12. Add 8µL of formic acid to each tube and mix well. Total sample volume is ~56µl. 16O- and 18O-labeled samples are ready to be mixed and analyzed by LC-MS.