eamag/protocols-with-steps huggingface dataset:
TITLE: Stable cell line generation via retrovirus
AUTHORS: Dan Tudorica

[DESCRIPTION]
Production of pseudotyped virus an subsequent transduction.

[STEPS]
1. 

Day 0: Thaw fresh cryovial of cells to
be transduced


Thaw ~ 1 week prior to planned transduction day, use
low passage number if possible. Aim for cells to be 80% confluent on day of transduction.
Recommend seeding four wells of a 12-well plate (100,000 cells/well) the day
before transduction (after collecting virus).


 


Day 1: HEK293T Transfection


     
Prepare 2 80% confluent 10 cm plates of
HEK293T for transfection


     
Prepare 3 mL of warm Opti-mem solution.
Add 10 µg of retro/lentiviral transfection plasmid, 10 µg VSV-G plasmid, and 10
µg pCMV-MLV (if retroviral)[DT1] [DT2] or 10 µg pCMV R8.74 (if lentiviral)


     
Add 90 µL of LT-1 reagent and swirl.


     
Incubate at room temperature for 15
minutes.


     
Add 1.5 mL dropwise to each 10 cm
HEK293T plate.


     
Incubate for 3 days in incubator before
harvesting viral particles. If a fluorescent marker was included, it should
begin to be visible on days 1,2 post transfection.


 


SAFETY: After
this point all media and plasticware is to be treated as potentially containing
viral particles. Decontaminate using 10% bleach for 10 minutes before
discarding as red bag waste. Rinse pipettes with 10% bleach before discarding
as biohazard waste.


 


Day 4: Harvest viral particles


     
Collect media from both HEK293T plates,
combine into one 20 mL portion in a 50 mL falcon tube. Bleach and discard used
HEK293T plate.


     
Spin 50 mL tube containing harvested
viral particles in Sorvall for 2 min at 2,000 rpm[DT3] [DT4] to clarify media and pellet stray cells


     
Aspirate off media without disturbing
pellet.


     
Add 6 mL Lenti-X concentrator solution
to 18 mL of recovered media (concentrator is 4x). Invert to mix.


     
Incubate in refrigerator 1 H – O/N[DT5] 


     
Spin 50 mL conical containing viral
particles in centrifuge at 1,500 rcf for 45 min


     
Remove 2 mL of supernatant from top of
pelleted solution and save.


     
Aspirate off remaining media to recover
viral pellet. Bleach excess media.


     
Resuspend retro/lenti pellet in 2 mL of
saved medium.


  
Titrate concentrated retro/lenti
solution into prepared 12-well containing target cells. Recommend
100-200-400-800 µL titration for 4 total attempts at transfection. Typically
400 or 800 uL will yield ~100% transfection efficiency


  
Return 12-well plate containing
transduced cells to incubator.


 


Day 5: Check on
transduction


     
May be able to see fluorescence today.


     
Swap media to fresh.


 


Day 6-7: Check on
transduction


 


Day 8-:


     
Allow cells to grow until almost
confluent on 12-well plate. Trypsinize and replate in a 6-well, allow to grow
until confluent, then replate in a 10 cm plate.


     
Split into two 10 cm plates, Freeze one
plate down into 4 cryovials the next day to save progress. Label with estimated
transduction efficiency if able to measure.


     
Take other plate and select with
antibiotics if able to/necessary, or clonally isolate populations if able
to/necessary.
TITLE: Marchantia genotyping (quick and dirty genomic DNA extraction)
AUTHORS: Eftychis Frangedakis, marta tomaselli, Marius Rebmann, Susana Sauret-Gueto

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">This protocol allows for quick and dirty genomic DNA extraction. It can easily be used for genotyping with PCR. The quality of the genomic DNA extracted is not suitable for any other application.</div><div class = "text-block"><span>It has been widely used in different plant species including Marchantia as in </span><a href="https://www.nature.com/articles/srep01532" style = "text-decoration:underline;color:blue;cursor:pointer;"><span style = ":;">https://www.nature.com/articles/srep01532</span></a></div></div>

[STEPS]
?. Take small pieces (3x3 mm) of thalli from individual plants and place in a 1.5 mL Eppendorf tube  (A in Figure).
?. Add 100 μl of genotyping buffer,: 100 mM Tris-HCl pH 9.5, 1M KCl, 10 mM EDTA (B in Figure).
?. Crush with an autoclaved micro-pestle (C in Figure).
?. Place the tube(s) at 80 °C for 10 min.
?. Add 380 μL of sterile water to each tube (D in Figure).
?. Use 5 μl aliquot of the extract as a template for PCR using preferably the KOD Hot start polymerase.
We found KOD to be more reliable amplyfing fragments from a crude genomic DNA extract such as the one used here.
?. Check PCR products on a 1.5% (w/v) agarose gel.
?.
TITLE: Light microscopic analysis of synaptic input to neurons in the rat major pelvic ganglion [keast-003]
AUTHORS: Janet Keast, Peregrine Osborne

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">This collection describes the procedures required to visualise and characterize synaptic boutons associated with functionally classified pelvic ganglion autonomic neurons of adult male and female Sprague-Dawley rats. This collection includes protocols for:</div><div class = "text-block">STAGE 1: Surgery to micro-inject fluorescent retrograde tracer dyes into one or more sites within the lower urinary tract</div><div class = "text-block">STAGE 2: Intracardiac perfusion with fixative to preserve neural tissues of interest</div><div class = "text-block">STAGE 3: Immunohistochemical labeling of thick cryosections of pelvic ganglia</div><div class = "text-block">STAGE 4: Confocal microscopy and image analysis of synaptic boutons associated with ganglion neurons.</div></div>

[STEPS]
TITLE: Slim Fast Keto Reviews: Is Keto Pills Really Works?
AUTHORS: Express Keto Pills  Reviews

[DESCRIPTION]
Slim Fast Keto Reviews: Is Keto Pills Really Works?

[STEPS]
SECTION: Slim Fast Keto Reviews: Is Keto Pills Really Works? Everyone wants to lose weight presto, numerous people are trying to get their dream body, and to some, it seems like a far- brought dream. Occasionally, losing too important weight too presto may be dangerous to your health.  For this reason, the world has embraced the ketogenic diet. Numerous people who are following the ketogenic diet are considering Slim Fast Keto. This supplement promises applaudable results, especially if you combine Slim Fast Keto mess plans with healthy exercises and a healthy diet.  In this Slim Fast Keto review, we will look at everything there&#x27;s to know about Slim Fast Keto Products. Where to buy Slim Fast Keto products? What are Slim Fast Keto snacks? Is it safe to take Slim Fast Keto mess plans? Are there Slim Fast Keto reviews from guests? Where to get Slim Fast Keto for trade online?  What Is Slim Fast Keto?  Slim Fast Keto, unlike other keto diets, isn&#x27;t a single product, and it&#x27;s five products under the marquee brand of Slim Fast Keto products. You can produce a whole keto diet plan when you put together the Slim Fast Keto products.  Slim Fast Keto is a clinically ratified keto diet plan.  The Slim Fast Keto products give its druggies a high fat, low- sugar, low carbohydrate ketogenic diet. The Slim Fast Keto uses ketosis to burn redundant fat in the body.  Slim Fast Keto diet plan provides numerous benefits to the druggies; some of the benefits include; hunger repression, rapid-fire fat- burning, and not having to give up your favorite foods.   Official Website:-https://freaknewswire.com/index.php/2022/06/03/slim-fast-keto-reviews-is-it-fake-or-trusted/ https://www.homify.in/projects/1152295/slim-fast-keto-reviews-is-keto-pills-really-work https://slim-fast-keto-reviews-is-keto-pills-really-works.jimdosite.com/ https://educatorpages.com/site/sofo62/pages/slim-fast-keto-reviews-is-keto-pills-really-works https://promosimple.com/ps/1fdf7/slim-fast-keto-reviews-is-keto-pills-really-works
1.
TITLE: Porcine Circovirus 3 (PCV3) Complete Genome Sequence PCR
AUTHORS: Chew Yee Tan

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">This protocol describes four separate PCR runs to amplify the complete genome sequence of Porcine circovirus 3 (PCV3) as four overlapping segments. </div></div>

[STEPS]
?. DNA extraction was performed using DNeasy Blood & Tissue Kit extraction kit (Qiagen, Germany) in accordance to manufacturer’s instructions. Samples involved include lung, inguinal lymph node, spleen, tonsil, kidney, mesenteric lymph node, heart, liver and brain tissues.
?. Four separate conventional PCR runs were performed to amplify the complete nucleotide sequence of PCV3 as four overlapping segments.
?. Primers, their respective cycling conditions and expected PCR product band size are tabulated below:
?. Type, concentration and volume of reagents used in the four separate PCR runs are the same: 12.5 μL of MyTaq™ Red Mix 2X master mix (Bioline, United Kingdom) and 1.0μM of each primer in a pair, in a 25 μL total PCR reaction volume.
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?. PCR products were stained using RedSafe™ nucleic acid staining solution (iNtRON Biotechnology, South Korea) and analysed by agarose electrophoresis using 2.5% agarose gel with GelPilot 100 bp Plus Ladder as marker (Qiagen, Germany).
TITLE: Conjugation of Keyhole limpet haemocynin to Peptide 254-274 of HIV gp-120 as immunogen.
AUTHORS: Angel  Justiz-Vaillant

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Chemical synthesis facilitates the generation of peptides which are difficult to express in bacteria. The fragment 254-274 of the human immunodeficiency virus (HIV) is critical for infectivity of the virus and induction of antibody neutralization (Ho et al, 1988). </div><div class = "text-block">Boudet et al, 1995 demonstrated that a synthetic peptide derived from the third variable domain of the HIV-1 gp-120 when used as immunogen was able to induce an antibody response to multiple (up to six) HIV strains. </div><div class = "text-block">Reference</div><div class = "text-block"><span>Ho DD, Kaplan JC, Rackauskas IE, Gurney ME. Second conserved domain of gp120 is important for HIV infectivity and antibody neutralization.</span><span style = "font-style:italic;">Science</span><span>. 1988;239(4843):1021-1023. doi:10.1126/science.2830667.</span></div><div class = "text-block"><span>Boudet F, Keller H, Kieny MP, Thèze J. Single peptide and anti-idiotype based immunizations can broaden the antibody response against the variable V3 domain of HIV-1 in mice.</span><span style = "font-style:italic;">Mol Immunol</span><span>. 1995;32(7):449-457. doi:10.1016/0161-5890(95)00007-2</span></div></div>

[STEPS]
?. The fragment 254-274 of gp 120 was conjugated by the glutaraldehyde method.
?. One mg of keyhole limpet hemocyanin (KLH) is dissolved in 2 ml 0.1 M borate buffer (1.24 g boric acid, 1.90 g sodium tetraborate, pH 10, in 500 mL deionized water).
?. In a 20 ml glass tube by gentle stirring 1 µmol of the HIV synthetic peptide and 0.2 mL 0.3% glutaraldehyde solution (ACS reagent grade, pH 5.5, Sigma-Aldrich) are slowly mixed at RT and left to stand for 2 hrs.
?. When a yellow coloration is observed this indicates that the conjugation process is successful.
?. To blocking the excess of glutaraldehyde, 0.26 ml of 1 M glycine (Sigma-Aldrich) is added.
?. The mixture is left for 31 min at RT.
?. The HIV-hemocynin conjugate is then dialyzed against 1.1 liter 0.1 M of borate buffer, pH 8.3 through the night at 4°C.
?. Then use borate buffer again to dialyze the preparation for 8 hrs at 4°C.
?. The dialysates is stored at 4°C.
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p>The <em>in vitro </em>α-glucosidase inhibitory assay was performed according to Dineshkumar et al. [8]. The α-glucosidase enzyme (2 U/mL) was premixed with 20 µL of plant aqueous extract at a concentration of 1,000 µg/mL and incubated for 5 min at 37 °C. Then 1 mM p-nitrophenyl gluco-pyranoside (pNPG) (20 µL) in 50 mM of phosphate buffer (pH 6.8) was added to initiate the reaction. The mixture was incubated at 37 °C for 20 min. The reaction was terminated by the addition of 50 µL of 1 mM sodium carbonate. The α-glucosidase activity was determined at 405 nm. The α-glucosidase inhibitory activity was calculated using the formula (Ac<sup>+</sup>) – (Ac<sup>-</sup>) – (As - Ab)/(Ac<sup>+</sup>) – (Ac<sup>-</sup>)× 100, where Ac<sup>+</sup>, Ac<sup>-</sup>, As, Ab are defined as the absorbance (405 nm) of 100% enzyme activity (only solvent with enzyme), 0% enzyme activity (only solvent without enzyme), test sample (with enzyme), and a blank (a test sample without enzyme), respectively. Reagents to evaluate anti-diabetic activity were of analytical grade and purchased from Sigma-Aldrich. For anti-diabetic activity, acarbose (anti-diabetic drug) was used as control.</p>
<p> </p>

[STEPS]
TITLE: Operating an OT-2 for COVID-19 testing
AUTHORS: Max Marrone

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

[STEPS]
?. Open the Opentrons App.
?. Click Start run. The OT-2 will home its motors and then begin the protocol.
Do not click Start run more than once. If you do, a known bug will make the OT-2 run the protocol back-to-back.
If something goes wrong and you need to abort the protocol:Shut down the OT-2 with the power switch on its back left side.Turn the OT-2 back on. Wait a couple of minutes for the pipettes to rise.Manually remove any tips attached to the pipettes. (This ensures that the pipettes will not aspirate liquid into themselves when they home.)Reconnect to the OT-2 in the Opentrons App. Click the Home button to move the gantry out of the way so you can access the labware on the deck.
?. Ensure you are connected to the robot. In the Robots tab, you can try flipping the robot's lights on and off to test the connection.
?. Go to the Run tab.Double-check the name at the top to make sure the correct protocol is uploaded.
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<div>
<div>
<div>
<p>This protocol is from 'Flower Ca<sup>2+</sup> channel in CME and ADBE' of Yao CK et al.</p>
<div>
<div>
<p> </p>
<p>Please see the manuscript for the full method details.</p>
</div>
</div>
</div>
</div>
</div>

[BEFORE_START]
<p>You'll need: </p>
<p> </p>
<p><strong>chamber (Warner instruments RC-25F) with two parallel platinum wires separated by 5 mm</strong></p>
<p> </p>
<p> </p>
<p><strong>imaging buffer:</strong></p>
<ul>
<li>136 mM NaCl</li>
<li>2.5 mM KCl</li>
<li>2 mM CaCl<sub>2</sub></li>
<li>1.3 mM MgCl<sub>2</sub></li>
<li>10 mM glucose</li>
<li>10 mM HEPES [pH 7.4]</li>
<li>10 mM CNQX</li>
<li>50 mM AP-5</li>
</ul>
<p> </p>
<p><strong>acidic buffer:</strong></p>
<ul>
<li>136 mM NaCl</li>
<li>2.5 mM KCl</li>
<li>2 mM CaCl<sub>2</sub></li>
<li>1.3 mM MgCl<sub>2</sub></li>
<li>10 mM glucose</li>
<li>10 mM 2-[N-morpholino] ethane sulphonic acid [pH 5.5]</li>
<li>10 mM CNQX</li>
<li>50 mM AP-5</li>
</ul>
<p>Acidic buffer was prepared by replacing HEPES in the imaging buffer with 2-[N-morpholino] ethane sulphonic acid [<a href="#_ENREF_47" target="_blank">47</a>,<a href="#_ENREF_48" target="_blank">48</a>]. </p>
<p> </p>
<p><strong>imaging solution:</strong></p>
<ul>
<li>144 mM NaCl</li>
<li>2.5 mM KCl</li>
<li>2.5 mM CaCl<sub>2</sub></li>
<li>2.5 mM MgCl<sub>2</sub></li>
<li>10 mM HEPES (pH 7.5)</li>
<li>10 mM CNQX</li>
<li>50 mM AP-5)</li>
</ul>

[GUIDELINES]
<p>Experiments were performed at either room or physiological temperatures controlled by Warner temperature controller (TC-344B).</p>
<p> </p>
<p> </p>
<p> </p>

[STEPS]
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?.
TITLE: Growing bacteria in Superbroth (“thick food”)
AUTHORS: Cristian Riccio

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><span>This protocol shows you how to prepare "thick food" to grow </span><span style = "font-style:italic;">C. elegans</span><span> at higher densities.</span></div></div>

[STEPS]
?. [Prepare Super Broth]
Add 1 bottle (100 ml) K-orthophosphates to each bottle of Super Broth (900 ml)
?. [Prepare starter culture]
Add 30 ml of Super Broth (plus K-orthophosphates) to a 100 ml flask. Inoculate Escherichia coli HB101 (also works with E. coli OP50, Acinetobacter schindleri, Bacillus pumilus and Pseudomonas fragi although not all these bacteria grow at 37 C) from plate in cold room. Use sterile flask and work under sterile conditions.Incubate for several (8 hours) hours at 37 °C, shaking (200 rpm). This culture can also be left to grow overnight.
?. [Prepare overnight cultures]
Add 1 L Super Broth (plus K-orthophosphates) to a 2.5 L sterile flask.Add ca. 5 ml starter culture (check that bacteria have grown!) to each 1L medium.Incubate over night at 37 °C, shaking (180-200 rpm). You may need to book an incubator to accommodate several flaks.
?. [Prepare the thick food]
Transfer cultures to 1000 ml plastic bottles for centrifugation. Use the large SorvallRC 3B Plus centrifuge, rotor H-6000.
?. [Prepare the thick food]
Spin cultures at 4000 rpm, 20 minutes, 4 °C.
?. [Prepare the thick food]
Discard supernatant and re-suspend pellet in 25-30 ml of H2O (sterile). Or b-broth.Work under sterile conditions to avoid contamination of bacteria.
?. [Prepare the thick food]
Seed culture using electronic pipette on low speed by dropping one droplet at atime, 2 ml in total on each of 14mm plate. The rationale behind this is thatworms like the edges of a bacterial lawn and seeding droplets increases thetotal length of edges.
?. [Prepare the thick food]
A 14mmplate can grow up to 80,000 animals.
?. [Prepare the thick food]
The bacteria can be frozen at -20 C for later seeding. A postdoc in the lab says to freeze resuspended bacterial while another postdoc says to freeze pellet. I personally froze the resuspended bacteria so that I can seed them immediately after defrosting.
?. [Prepare Super Broth]
TITLE: Indications and prescriptions of penicillins in a population of Colombia: A cross-sectional study
AUTHORS: Jorge Machado Alba

[DESCRIPTION]
Introduction: Inappropriate
use of antibiotics plays a key role in increasing bacterial resistance, which
is a global problem that has increased over time.
 
Objective: To
determine the prescription patterns and indications for the use of penicillins
in a group of patients from Colombia.
 
Methods: This study was a
cross-sectional study on the use of penicillins in outpatients. Approved and
unapproved indications were determined from records of the Food and Drug
Administration (FDA) of the United States and the National Institute of Food
and Drug Surveillance (INVIMA) of Colombia. Descriptive and multivariate analyses
were performed.
 
Results: A total of 137,070
patients were identified; the average age was 35.8 ± 23.5 years, and 56.2% were
women. Amoxicillin (73.4%), dicloxacillin (11.7%) and sultamicillin (6.0%) were
the most commonly prescribed penicillins, mainly for upper respiratory tract
infections (43.0%). In 68.9% of cases, penicillins were used for approved
indications, especially to treat Helicobacter
pylori (17.3%). In 31.1% of cases, penicillin prescriptions were used for
unapproved indications (e.g., acute rhinopharyngitis [8.1%]). Patients under 18
years of age (OR: 1.87; 95% CI: 1.78-1.96), those from municipalities (OR:
1.51; 95% CI: 1.44-1.57), those with previous antibiotic use (OR: 2.49; 95% CI:
2.26-2.74), those with skin and soft tissue infections (OR: 2.82; 95% CI:
2.57-3.09), those with lower respiratory tract infections (OR: 2.02; 95% CI:
1.89-2.16), those managed with general medicine (OR: 1.37; 95% CI: 1.25-1.51),
and those treated with dicloxacillin (OR: 2, 84; 95% CI: 2.07-3.89) were more
likely to be prescribed penicillins for unapproved indications.
 
Conclusions:
Amoxicillin was the most widely used penicillin in outpatients. Penicillins
were frequently used for unapproved indications not recommended by clinical
practice guidelines.
 

[STEPS]
TITLE: Genotyping protocol for mice
AUTHORS: vanessa promes

[DESCRIPTION]
This protocol describes the general method to genotype the mice.

[STEPS]
SECTION: DNA Extraction
1. Obtain tail sample from mice.
SECTION: DNA Extraction
2. Add 100 µL of NaOH into tail sample and place in hot plate at 100C for 10 min. Vortex sample and place in hot plate once more for 10 min. 

SECTION: DNA Extraction
3. Add 50 µL of 1M Tris pH 8.0 to neutralize extraction

SECTION: DNA Extraction
4. Spin for 20,000xg for 10 min

SECTION: DNA Extraction
5. Use 2µL in PCR reaction.
SECTION: Preparing PCR reaction
6. Obtain a 1.5ml microcentrifuge tube and add 10ul 2x DreamTaq Green Master mix. 2ul forward and reverse primers, 2µL DNA lysate and Nuclease-free water up to 20µL
SECTION: Preparing PCR reaction
7. Thermocycling parameters:
95oC 5min
95oC 30sec| x 30 cycles
60oC 30sec|
72oC 1min|
72oC 2min
4oC hold
SECTION: Run DNA Sample
8. In 2% agarose in TAE buffer add 10ul of the sample and run for around 30min
TITLE: Miniprep (NEB Monarch)
AUTHORS: Brian    Teague

[DESCRIPTION]
We transformed E. coli bacteria with our plasmid in order to make more of it -- to use the bacteria as highly accurate DNA copiers. Now that we've grown a bunch of E. coli, we need to get the plasmid DNA back out. That's the point of a miniprep.

  


[GUIDELINES]
It is easy to get "into a rhythm" with this protocol and miss or mis-read important steps. Follow the directions very carefully -- there are lots of buffers and centrifugation stepsS

[STEPS]
SECTION: Harvest the E.coli culture
1. Transfer 1.5 mLof bacterial culture to a microcentrifuge tube. Centrifuge 16000 x g, 30 s

SECTION: Harvest the E.coli culture
2. Remove the supernatant using a micropipettor and discard in the biological waste container. Try to get as much as you can without disturbing the pellet.
SECTION: Harvest the E.coli culture
3. Transfer another 1.5 ml of bacterial culture to the same microcentrifuge tube. Centrifuge16000 x g, 30 s

SECTION: Harvest the E.coli culture
4. Remove the supernatant using a micropipettor and discard in the biological waste container. Try to get as much as you can without disturbing the pellet.
SECTION: Lyse the E.coli cells
5. Add 200 µL of resuspension buffer. Vortex to resuspend the pellet. Make sure it is completely resuspended -- there should be no visible clumps.

SECTION: Lyse the E.coli cells
6. Add 200 µL lysis buffer. Invert the tube 4-6 times to mix and incubate at Room temperaturefor 1 min. Do not vortex.

SECTION: Lyse the E.coli cells
7. Add 400 µL of neutralization buffer and gently invert the tube until neutralized. Sample is neutralized when the color is uniformly yellow. Do not vortex.

SECTION: Lyse the E.coli cells
8. Incubate at Room temperature for 2 min

SECTION: Lyse the E.coli cells
9. Centrifuge lysate 16000 x g, 5 min

SECTION: Bind and Wash
10. Carefully transfer the supernatant (the liquid at the top, not the pellet at the bottom) to the spin column. Centrifuge 16000 x g, 1 min. Discard the flow-through in the chemical waste container.

SECTION: Bind and Wash
11. Re-insert column in collection tube. Add 200 µL of wash buffer 1. Centrifuge 16000 x g, 1 min. Discard the flow-through in the chemical waste container.

SECTION: Bind and Wash
12. Add 400 µL of wash buffer 2. Centrifuge 16000 x g, 1 min. Discard the flow-through in the chemical waste container.

SECTION: Bind and Wash
13. Replace the spin column in the (now empty) collection tube. Centrifuge 16000 x g, 1 min 

 

SECTION: Elute
14. Transfer the column to a clean 1.7 ml microcentrifuge tube. Be careful: make sure the tip of the column doesn't come into contact with the flow-through.
SECTION: Elute
15. Add 30 µL of elution buffer to the center of the silica matrix. Wait 1 min, then centrifuge 16000 x g, 1 min

SECTION: Elute
16. Measure the concentration and purity of your DNA on the NanoDrop.
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p>This protocol describes protein extraction steps from mouse aorta or vascular smooth muscle cell sample for Western blot. The lysis buffer use is a SMAD lysys buffer adopted from Beaufort et al (2014).</p>

[STEPS]
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?.