eamag/protocols-with-steps huggingface dataset:
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<div title="Page 1">
<div>
<div>
<p>Endo H cleaves Asparagine-linked hybrid or high mannose oligosaccharides, but not complex oligosaccharides. It cleaves between the two N-acetylglucosamine residues in the diacetylchitobiose core of the oligosaccharide, generating a truncated sugar molecule with one N-acetylglucosamine residue remaining on the asparagine. In contrast, PNGase F removes the oligosaccharide intact. Detergent and heat denaturation may increase the rate of cleavage for some glycoproteins.</p>
</div>
</div>
</div>

[STEPS]
?. 
?. 
?. 
?.
TITLE: FUNDIS ONT Sequencing IT/Compute Pop!_OS 22.04 Setup
AUTHORS: Stephen Douglas Russell, Harte Singer

[DESCRIPTION]
These are the steps to setting up a Linux system running Ubuntu for ONT Nanopore sequencing, basecalling, and consensus-building of fungal amplicon sequences. Adapted from dx.doi.org/10.17504/protocols.io.14egn7kzmv5d/v3

Includes an R-based run QC script

[STEPS]
SECTION: Preparing a new CPU for MinION Sequencing
1. Ideally you will be running a linux machine with a Ubuntu distro with an NVIDIA graphics card with at least 8GB of ram. (I am running a 4070Ti) You should have at least 32GB of system ram and a new processor. 

 


Minimum IT requirements for MinION from ONT:    
SECTION: Preparing a new CPU for MinION Sequencing
2. 

Install CUDA toolkit - https://developer.nvidia.com/cuda-toolkit :
 

SECTION: Preparing a new CPU for MinION Sequencing
3. Install Boost

 

SECTION: Preparing a new CPU for MinION Sequencing
4. Install MinKNOW from https://community.nanoporetech.com/downloads?from=support

Specifically download the GPU version of the software with the following name. It is a .deb file and your computer will install it when you double-click it

MinKNOW Software for the MinION Mk1B and the PromethION 2 Solo (P2 Solo) Devices

SECTION: Preparing a new CPU for MinION Sequencing
5. Install Guppy from the "Archived Software" section at the bottom of this page https://community.nanoporetech.com/downloads?from=support

Guppy v6.5.7

Make sure you install the Ubuntu 20 GPU version, just double click the installer and it should work. 








SECTION: Preparing a new CPU for MinION Sequencing
6. Per this document: https://denbi-nanopore-training-course.readthedocs.io/en/latest/read_qc/MinionQC.html
Install R: https://cran.r-project.org/
Install MinionQC: https://github.com/roblanf/minion_qc

Install R:

 


 
Open an R terminal window for the following command
 

SECTION: Preparing a new CPU for MinION Sequencing
7. Install Bioconductor:

In an R command window:

 

SECTION: Preparing a new CPU for MinION Sequencing
8. Install Anaconda: 
from: https://www.digitalocean.com/community/tutorials/how-to-install-the-anaconda-python-distribution-on-ubuntu-22-04


 

 

 

 

 

 

 

 

 

 



 


 



SECTION: Preparing a new CPU for MinION Sequencing
9. Install NGSpeciesID: https://github.com/ksahlin/NGSpeciesID


 

 

 

 

 

 

SECTION: Preparing a new CPU for MinION Sequencing
10. You should now be ready to begin sequencing runs.
SECTION: Installing Dorado
11. We will eventually be moving to Dorado basecalling. 

Installing Dorado:
TITLE: Passaging of organoids (organoids split into cell clusters)
AUTHORS: Gabriela Vallejo Flores, Annika Fendler

[DESCRIPTION]

The goal of this experiment is to split the organoids into cell clusters and induce organoids formation and organoids expansion.

[BEFORE_START]
Prepare the organoids medium one day before organoids splitting
Pre-warm seeding plate
work with 1% BSA/PBS coated tubes and pipettes
use pre-cooled pipette tips for matrigel
Thaw the matrigel 1hr before starting the experiment and prepare the multiwell plate for each protocol; 
 




[STEPS]
SECTION: Organoids Splitting
2. Remove the supernatan from the well and add 1 mL   to release the organoids from the matrigel, 400 x g, 5 min, 4 °C, and aspirate the supernatant and Matrigel 
above the pellet.  Note:  For the minoli protocol "organoids grow in cell suspention", collect the supernatant and do the centrifugation. 
SECTION: Organoids Splitting
3. Resuspend the organoids in 500 µL (double the amount of Tryple Express when the matrigel is still visible after removing the supernatant, and incubate for15 min  with frequent perturbation by pipetting, check organoid dispersion under the bright-field microscope every 5 min. 

Quantity of TripLE exprees per well, Table 1
 

SECTION: Organoids Splitting
4. Add  1 mL  to dilute the TripLE express and incativate the enzyme,  400 x g, 5 min, 4 °C, and aspirate the supernatant.
SECTION: Organoids Splitting
5. Homogenize the pellet in appropriate volume of Matrigel and pipet into a pre warmed plate: prewarme
 

SECTION: Cell seeding Lee
9. Add the Matrigel per 20x10^3 cells, per well and incubate the plate during 15 min at 37°C, 5%CO2 ;
Add the matrigel, as described in the Table 4.

 

SECTION: Organoids Splitting
1. 
 

SECTION: Cell seeding Mullenders
6. Add matrigel in pre wamed plate, as described in Table



 

SECTION: Cell seeding Mullenders
7.  Flip the 48,24 or 6 wells plate and incubate 15 min at 37°C 5%CO2.
SECTION: Cell seeding Mullenders
8. Add 250µl, 500µl or 2ml of medium per well. 
SECTION: Cell seeding Lee
10. Add 250µl,  500µl or 2ml of medium per well.
TITLE: Improve 
AUTHORS: Hung Yu Chen, Huan Jui Chang

[STEPS]
?. [In vitro protIn vitro protein synthesis]
Add the reagents(on ice) for synthesis of amilCP into 15 wells of the 384-well plate. Pipette well before adding reagents into the wells.banana amilCP  ABCD1OrderLocationReagentAmount21Upper-leftDNase/ RNase free watertill 5 μL32Upper-leftSolution A2 μL43Lower-rightSolution B1.5 μL54Lower-rightRNase inhibitor0.2 μL65Lower-leftbanana amilCP50 ng7Total5 μLbanana trigger + banana amilCP  ABCD1OrderLocationReagentAmount21Upper-leftDNase/ RNase free watertill 5 μL32Upper-leftSolution A2 μL43Lower-rightSolution B1.5 μL54Lower-rightRNase inhibitor0.2 μL65Lower-leftbanana amilCP50 ng76Lower-leftbanana trigger50 ng8Total5 μLT7 Strong Promoter + amilCP  ABCD1OrderLocationReagentAmount21Upper-leftDNase/ RNase free watertill 5μL32Upper-leftSolution A2 μL43Lower-rightSolution B1.5 μL54Lower-rightRNase inhibitor0.2 μL65Lower-leftT7 Strong Promoter amilCP50 ng7Total10 μL
ABCD1OrderLocationReagentAmount21Upper-leftDNase/ RNase free watertill 5 μL32Upper-leftSolution A2 μL43Lower-rightSolution B1.5 μL54Lower-rightRNase inhibitor0.2 μL65Lower-leftbanana amilCP50 ng7Total5 μL
ABCD1OrderLocationReagentAmount21Upper-leftDNase/ RNase free watertill 5 μL32Upper-leftSolution A2 μL43Lower-rightSolution B1.5 μL54Lower-rightRNase inhibitor0.2 μL65Lower-leftbanana amilCP50 ng76Lower-leftbanana trigger50 ng8Total5 μL
ABCD1OrderLocationReagentAmount21Upper-leftDNase/ RNase free watertill 5μL32Upper-leftSolution A2 μL43Lower-rightSolution B1.5 μL54Lower-rightRNase inhibitor0.2 μL65Lower-leftT7 Strong Promoter amilCP50 ng7Total10 μL
Do triple repeat
?. [In vitro protIn vitro protein synthesis]
Seal the plate with microseal (on ice)
?. [In vitro protIn vitro protein synthesis]
Centrifuge (4000rpm, 0.01 (1 min), 4°C)
?. [Setup plate reader]
Temperature: Setpoint 37°CShake: Orbital for 1:00 (1 min)Start Kinetic [ Run 6:00:00 (6 hour), Internal 0:05:00 (5 min)]Read: (A) 588 nmEnd Kinetic
?. [Incubation]
Put the plate into plate reader.Incubate the mixture and measure the fluorescence in plate reader follow Setup 1 for 06:00:00
?. [Measuring]
Measure the fluorescence excitation and emission intensity of the parts.
TITLE: Fluorescent immunochemistry
AUTHORS: Corentin Cras-Méneur

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">General fluorescent immunostaining protocol for paraffin sections</div></div>

[STEPS]
?. [Deparaffinizing]
- - - Ethanol 100°   - Ethanol 95°  - Ethanol 75°
?. [Antigen Retrieval]
Microwave for 3 x  with microwaving solution (kept @ )Let it cool until
4 °C
on ice
0 Room temperature
?. [Permeabilization]
Permeabilization with TBS 1x;  Triton 100 (20 min @ )
0 Room temperature
?. [Blocking]
Blocking with the Blocking Solution
?. [Primary antibody]
Anti Insulin (Guinea Pig) 1/800Anti Ki67 (Rabbit) 1/200( @)
4 °C
?. [Wash]
Wash with TBS 1 x; 0.1% Tween, rocking (3 x )
?. [Secondary Antibody]
Anti Guinea Pig-FITC 1/800Anti Rabbit-Cy3 1/200( @ )
[minimum]
4 Room temperature
?. [Wash]
Wash with TBS 1 x; 0.1% Tween, rocking (3 x )
?. [Final wash]
Wash with TBS 1 x rocking ()
?. [Mount]
Coverslip with antifading agent mounting media with or without DAPIUse Anti-fading Mounting Media
?. [Antigen Retrieval]
Microwaving solution:9 mL Citric Acid 0.01 M
							  .justify:after {
							    content: "";
							    display:inline-block;
							    width: 100%;
							  }
							41 mL Sodium Citrate 0.01 M
							  .justify:after {
							    content: "";
							    display:inline-block;
							    width: 100%;
							  }
							450 mL H2O
TITLE: Quick and dirty sequencing microbial genome extraction v1
AUTHORS: Sung won Lim, Sebastian Cocioba

[DESCRIPTION]
Quick and dirty microbial genome extraction protocol using Edwards buffer. 

I've been using this method for in-lab Archaeal evolution study for the last four years or so and it's been working fantastically well, especially for short read sequencing. 

Certain microbes (such as Deinococcus) can be difficult to process properly using just this protocol - standard caveats and fixes tend to improve the output, such as adding initial lysozyme incubation step in the very beginning, sometimes combined with a freeze-thaw cycle for especially difficult samples (I'll update and upload a separate version of this protocol for difficult microbes in the future).

This particular protocol is aimed at getting as much intact, long-read capable DNA out from a microbe as cheaply as possible, as shown in the included gel picture. 

Originally written up for my lab note at https://naturepoker.wordpress.com/2023/02/26/halobacteria-mutant-sequencing-3-years-in/

Ideation and initial testing performed with Sebastian S. Cocioba at Binomica Labs for ONT Deinococcus radiophilus genome sequencing project, using RAD004 rapid sequencing kit. 

https://www.ncbi.nlm.nih.gov/assembly/GCF_020889625.1

[STEPS]
1. Spin down 1 ml of sample for 1 minute at max speed and decant
2. Resuspend vigorously with 100ul Edward's buffer
3. Transfer carefully to PCR tube - mixture will be viscous
4. Add 2ul of RNase A and mix vigorously, vortex for 10 seconds
5. Incubate at 37C for 15 minutes
6. Add 2ul of Proteinase K and mix vigorously, vortex for 10 seconds
7. Incubate at 55C for 1 hour, and deactivate via incubation at 95C for 10 minutes
8. Transfer to 1.5ml eppendorf tube
9. Add 10% 3M (pH 5.4) sodium acetate, and 1:1 volume of 100% isopropyl alcohol
10. Invert tube 10 times - precipitates should begin to form
11. Spin down at max speed for 5 minutes
12. Decant the supernatant carefully
13. Add 1ml of 70% EtOH and resuspend the pellet
14. Spin down at max speed for 5 minutes
15. Repeat 70% EtOH resuspension and washing step at least 2 more times
16. Decant completely and dry the pellet for 5 minutes - do not let the pellet overdry
17. Resuspend in storage buffer of choice or dH2O to wanted volume 
18. Incubate in a 37C shaker for 1 hour - the extract is likely to be not dissolved fully
19. Incubate in 4C overnight - depending on yield, the extract will be extremely viscous
20. Check extraction quality using both standard gel electrophoresis and UV-Vis method such as Nanodrop.
TITLE: SureSelect XT HS2 DNA to prepare libraries for single-cell Whole Genome Sequencing (scWGS) after single-cell Whole Genome Amplification (scWGA)
AUTHORS: Ester Kalef-Ezra, Ben Harvey, Katherine Roper, Christos Proukakis

[DESCRIPTION]
We adapted the SureSelect XT HS2 DNA protocol to prepare libraries using as input material samples after single-cell Whole Genome Amplification (scWGA), instead of genomic DNA samples, for library preparation for Illumina single-cell Whole Genome Sequencing (scWGS). This protocol can be employed either manually (Section 2: Option A) or in combination with automation using the Bravo Automated Liquid Handling Platform (Section 2: Option B).

[GUIDELINES]
We have used this protocol for DNA products after single-cell Whole Genome Amplification (scWGA) by either PicoPLEX (Takara), ResolveDNA (BioSkryB) and droplet MDA (Samplix). We recommend to bead purify the PicoPLEX and ResolveDNA products prior library preparation. In all cases, the amplicon quantity and size should be assessed using Qubit and TapeStation or other methods prior to SureSelect XT HS2 DNA library preparation.


Bravo error/notification messages

Two error/notification messages are encountered when VWorks Bravo software is initializing: 

1. If you encounter the G-axis error message shown below, select Ignore and Continue, leaving device in current state.
  2. If you encounter the W-axis error message shown below, select Retry. 
 


[STEPS]
TITLE: Streptavidin dot blot with aptamers
AUTHORS: Manuela de las Casas

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">The aim of this protocol is to check the efficiency of the detection system that will be present in the strips. The result expected is a good binding between the streptavidin and the aptamers, great hybridization between both aptamers and a clear visible result when yielding with peroxidase. </div></div>

[STEPS]
?. [Preparing the nitrocellulose strip]
Set the hot plate to 100ºC or at least warm enough to melt the wax. Cut a small amount from one of the wax pencils and place it  on a Petri dish. Set the Petri dish on the hot plate and wait for the wax to melt. Cut a strip from the nitrocellulose sheet with the desired size.Once the wax is melt, grab a 200μL pipette tip from the pointy side and place the broad end in the wax. Check the whole circle contains enough wax and carefully place it on the nitrocellulose strip. Apply a little pressure and remove the pipette tip. You should be able to see the wax circle on the strip.Cover the nitrocellulose strip with paper (one sheet on top, another underneath). Place the sandwich on the hot place with a Petri dish on top (this is just to apply some pressure, any temperature-resistant flat object is valid here). Allow the strip to warm for 5 minutes. This will allow the wax to penetrate all through the strip.Remove the strip from the hot plate and let it cool down again. You should see a circle of wax but the center must be clear, otherwise the protein will not have anywhere to bind. If the circle is too thick, consider repeating these steps.
?. [Adding the streptavidin and blocking the membrane]
Pipette 5 μL of Streptavidin. If the protein is too diluted, you might not see a staining with the Ponceau. We ended using the protein suspended in PBS but without any dilutions. Allow it to dry completely. Room temperature is adequate. Once the membrane is completely dry, pipette 5 μL of the blocking solution (BSA, powder milk and Tween 20) to block the empty spaces left in the membrane. Allow it to dry completely again.
?. [Hybridization of the aptamers]
TITLE: Dual Luciferase Assay 
AUTHORS: vahedilaboratory  

[DESCRIPTION]
Dual luciferase assay WIP

[STEPS]
3.  Cell Lysing and Luciferase Assay
Remove media from each well and wash with PBS; remove the PBS
Add 50uL trypsin to each well and coat every cell
Leave in incubator for 3 minutes
Pipette all cells off of plate and add cells to 1.5mL tubes 
Spin down cells at 1,500RPM 4C for 5 minutes to pellet cells 
Remove supernatant 
Re suspend cells in 50uL of PLB 1X Passive Lysis Buffer (PLB)
Incubate lysates on ice for at least five minutes
Vortex each tube for 10-15 seconds
Incubate for 5-10 more minutes on ice
Centrifuge the tubes at 12,000 rcf at 4C for two minutes
Transfer the supernatant of each tube to a new labeled tube.
Either store the lysates at -80C or continue to the luciferase assay
1. Cell culture 
If you are reading this, I am assuming that you already have 100,000 cells plated in each well of a 24-well TC plate (or something of equal measure)
Let them rest overnight because tomorrow you will transfect them 
4. The Luciferase Assay Itself
Set up a GloMax luminometer to measure for 10 seconds
For each sample you want to measure, add 20 uL of LARII and 20 uL lysate to a microfuge tube
Vortex the tube briefly
Put the tube in the slot in the luminometer, close the lid, and measure
Usually measure each sample two times
2. Transfection (Transient) of Plasmid
Dual luciferase assays work by expressing and measuring two luciferase plasmids: Firefly Luciferase and Renilla luciferase, which helps normalize transfection efficiencies of the plasmids 
Assemble the following mixture for a 24 well plate: 

2uL of FuGENE
1uL plasmid A (200ng)
1uL plasmid B (200ng)
50uL Optimem

Let sit for max 15 minutes 
Add dropwise to the cells 
Leave for 24 hours
TITLE: LyGo cloning
AUTHORS: Kristoffer Bach Falkenberg, Cristina Hernandez Rollan, Maja Rennig, Andreas Birk Bertelsen, Morten Norholm

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Lytic polysaccharide monooxygenses (LPMOs) are enzymes that play a critical role in breaking the chemical bonds of the most abundant polymers found in recalcitrant biomass, such as cellulose and chitin. LyGo cloning (Lytic Polysaccharide Monooxygenase Golden Gate cloning) is a versatile heterologous expression platform for LPMOs, which is compatible with cloning both PCR products and synthetic gene fragments with a simple 15-minute assembly step. The method allows for parallel construction of multiple expression vectors, enabling exploration of several expression strategies. The open-source LyGo collection consists of vectors for some of the most relevant model organisms used for protein production in both academic and industrial settings.</div><div class = "text-block">This protocol describes how to clone LyGo fragments into LyGo vectors.</div></div>

[STEPS]
?. [Mix reactions]
Prepare a reaction mix in the following proportions:  AB1ComponentVolume (μL)2LyGo vector13LyGo fragment34FastDigest SapI0.5510x T4 ligase1610x FastDigest buffer1710x T4 buffer18MQ2.59Total10
AB1ComponentVolume (μL)2LyGo vector13LyGo fragment34FastDigest SapI0.5510x T4 ligase1610x FastDigest buffer1710x T4 buffer18MQ2.59Total10
The exact volume of LyGo vector and LyGo fragment depends on their kb sizes and DNA concentrations. The optimal molar ratio of vector:fragment is between 1:3 and 1:6, so dilutions/additions should be made accordingly. The NEB Ligation Calculator (https://nebiocalculator.neb.com/#!/ligation) is a useful tool to perform these calculations.
?. [Reaction]
Incubate the reactions for at least  at .
22 Room temperature
?. [Reaction]
The reactions are now ready to be transformed directly into competent E. coli cells, using established protocols e.g.
{"blocks":[{"key":"9d6re","text":"This describes a method to transform a plasmid into homemade DH5\u03b1 cells.","type":"unstyled","depth":0,"inlineStyleRanges":[],"entityRanges":[],"data":[]}],"entityMap":[]}
In our experience, it is helpful to transform all   of the reactions, although this might depend on the ligation and transformation efficiencies.
Subsequent construct verification should also be performed (e.g. PCR, digestion and/or sequencing), before transforming the expression vectors into the expression host.
TITLE: Nested PCR amplification of Salmonella Typhi from extracted wastewater concentrate
AUTHORS: Shannon Fitz, Alex Shaw, michael Owusu, Dilip Abraham, Anton Spadar

[DESCRIPTION]
This standard operating procedure describes nested PCR protocols for the generation of amplicons from V. cholerae and S. typhi.   

PCR primers have been sourced from the literature or designed using Geneious, with primer pooling tested to minimise primer-primer interactions. Whilst the first round of PCR may be sufficient alone for detection (and will yield larger amplicons) a second round of PCR has been included to increase sensitivity.  

[STEPS]
SECTION: Primer preparation step
2. Workstation Preparation

Clean the PCR areas before setting up the reactions; clean room for the master mix and an area for the addition of the template. 
Clean the working area and pipettors with an approved DNAse inhibitor solution.
Clean the Class II biosafety cabinet and pipettes with 70% ethanol and wipe with a paper towel.
Place a disposal Dispo-safe “sweetie” jar or bio-bin on the bench. 

SECTION: Primer preparation step
1.  
 
Primer preparation 
 
Reconstitute primers to 100 µM using nuclease-free water 
Create working stocks of 10 µM using nuclease-free water 
Create 10 µM  primer pools G and B by mixing together 10 µL of each primer marked as being part of the pool. Scale up as needed.   
SECTION: Primer preparation step
3. Sample Retrieval

Place a rack inside a polystyrene sample box for sample transfer.  
Remove tube containing the required samples from the -80 °C freezer.  
Close freezer and polystyrene box. 
Allow the samples to thaw on ice. 
SECTION: Primer preparation step
4. Round 1 PCR reaction

Prepare the following Master mix in a 1.5ml Eppendorf Lobind tube: 
 

Briefly vortex and centrifuge down the master mix and aliquot 20 µL into each PCR tube. 
Add 5 µL of extracted DNA from each sample to a tube, mix well by pipette and label accordingly. 
Briefly centrifuge down the PCR mixes and store on ice. 

SECTION: Primer preparation step
5. Amplify using the following cycling conditions: 

 

 
SECTION: Primer preparation step
6. Round 2 PCR reaction

 Prepare the following Master mix in a 1.5ml Eppendorf Lobind tube: 
 
Briefly vortex and centrifuge down the master mix and aliquot 20 µL into each PCR tube. 
Add 5 µL of extracted DNA from each sample to a tube, mix well by pipette and label accordingly. 
Briefly centrifuge down the PCR mixes and store on ice. 
SECTION: Primer preparation step
7. Amplify using the following cycling conditions: 
 
 
 
SECTION: Primer preparation step
8. Gel electrophoresis bands
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p>This protocol outlines our preparation of single-cell suspension from surgically acquired fresh human adult brain tissue. This method is adapted from Habib et al<sup>1</sup>.</p>
<p> </p>
<p>1          Habib, N.<em> et al.</em> Massively parallel single-nucleus RNA-seq with DroNc-seq. <em>Nat Methods</em> <strong>14</strong>, 955-958, doi:10.1038/nmeth.4407 (2017).</p>

[STEPS]
?. 
?. 
?. 
?. 
?. 
?. 
?.
TITLE: Preparation of the adenylated oligonucleotide for mapping 3′OH RNA termini
AUTHORS: Matus Valach

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><span>Adapted from the protocol for oligonucleotide 5′ adenylation (E2610) of </span><span style = "font-style:italic;">New England Biolabs</span><span> (</span><span style = "font-style:italic;">NEB</span><span>).</span></div></div>

[STEPS]
?. Mix the following components (20 μL):  ABC1ComponentAmount [μL]Final concentration25′ phosphorylated DNA oligo [50 μM]410 μM310× 5′ DNA Adenylation buffer21×4ATP [1 mM]20.1 mM5Mth RNA ligase [50 pmol/µL]410 µM6RNase-free water (ddH2O)8
ABC1ComponentAmount [μL]Final concentration25′ phosphorylated DNA oligo [50 μM]410 μM310× 5′ DNA Adenylation buffer21×4ATP [1 mM]20.1 mM5Mth RNA ligase [50 pmol/µL]410 µM6RNase-free water (ddH2O)8
?. Incubate for 60 minutes at 65 °C.
?. Incubate for 5 minutes at 85 °C to inactivate the enzyme.
?. The adenylated oligo can be directly used for RNA ligation with T4 RNA ligase 2 (optimally with T4 RNA Ligase 2, truncated KQ (NEB #M0373)).
TITLE:  Analysis of the Derivatives of Sialic Acid in Pig Tissues
AUTHORS: Chon-Ho Yen

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Sialic acid is a generic term for the N- or O-substituted derivatives of neuraminic acid, a monosaccharide with a nine-carbon backbone. N-glycolylneuraminic acid (Neu5Gc or NGNA) is not synthesized in the human body, so most of the sialic acid in the body is N-acetylneuraminic acid (Neu5Ac or NANA), while other animals have two kinds of sialic acid (NANA and NGNA). There may be NGNA in biotech products manufactured from nonhuman animal source, and a detectable method is needed to determine the derivatives of sialic acid in the product. </div></div>

[STEPS]