eamag/protocols-with-steps huggingface dataset:
TITLE: WILLIS -HOBBS AGAR
AUTHORS: Roey Angel, Ana Lara-Rodriguez

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">For the diferential isolation of Clostridium sp. </div></div>

[STEPS]
?. For the preparation of 1L of media (50 to 66 petri dishes depending of depth) weigh:Final pH ( at 25°C) 7.0±0.2
[ Peptic Digest from Animal tissue]
[ Meat Extract]
[ NaCl]
[ Lactose]
[ Neutral Red]
[Agar]
[Egg yolk Emulsion]
?. Heat to boiling to dissolve the medium completely.
?. Sterilize by autoclaving at 15 lbs pressure  for .
121 °C
?. Cool to
50 °C
?. Add  sterile skimmed milk 10% solution.
120 mL
?. Aseptically add Egg Yolk Emulsion.
30 mL
?. Pour  or   of media in petri dishes depending on desired depth .
15 mL
20 mL
?. Let cool the open petri dishes under UV light for  to  .
?. Close petri dishes and keep at   until use. (2 weeks max)
4 °C
TITLE: Bivalent binding of a fully human IgG to the SARS-CoV-2 spike proteins reveals mechanisms of potent neutralization
AUTHORS: Bei Wang, Daniel Asarnow, Wen-Hsin Lee, Ching-Wen Huang, Bryan Faust, Patricia Miang Lon Ng, Eve Zi Xian Ngoh, Markus Bohn, David Bulkley, Andrés Pizzorno, Hwee Ching Tan, Chia-Yin Lee, Rabiatul Adawiyah Minhat, Olivier Terrier, Mun Kuen Soh, Frannie Jiuyi Teo, Yvonne Yee Chin Yeap, Yuanyu Hu, Shirley Gek Kheng Seah, Sebastian Maurer-Stroh, Laurent Renia, Brendon John Hanson, Manuel Rosa-Calatrava, Aashish Manglik, Yifan Cheng, Charles S. Craik, Cheng-I Wang

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><span style = "font-style:italic;font-weight:bold;">In vitro </span><span style = "font-weight:bold;">antibody selection against pathogens from naïve combinatorial libraries can yield various classes of antigen-specific binders that are distinct from those evolved from natural infection. Also, rapid neutralizing antibody discovery can be made possible by a strategy that selects for those interfering with pathogen and host interaction. Here we report the discovery of antibodies that neutralize SARS-CoV-2, the virus responsible for the COVID-19 pandemic, from a highly diverse naïve human Fab library. Lead antibody 5A6 blocks the receptor binding domain (RBD) of the viral spike from binding to the host receptor angiotensin converting enzyme 2 (ACE2), neutralizes SARS-CoV-2 infection of Vero E6 cells, and reduces viral replication in reconstituted human nasal and bronchial epithelium models. 5A6 has a high occupancy on the viral surface and exerts its neutralization activity via a bivalent binding mode to the tip of two neighbouring RBDs at the ACE2 interaction interface, one in the “up” and the other in the “down” position, explaining its superior neutralization capacity. Furthermore, 5A6 is insensitive to several spike mutations identified in clinical isolates, including the D614G mutant that has become dominant worldwide. Our results suggest that 5A6 could be an effective prophylactic and therapeutic treatment of COVID-19.</span><span></span></div></div>

[STEPS]
TITLE: gynecomastia surgery
AUTHORS: habedih  

[DESCRIPTION]
Gynecomastia is a reviewed condition portrayed by the development of the male bosom that influences a critical extent of the male populace. Plenty of fluctuating careful methodologies at present exists in the writing; consequently, this complete audit tried to break down careful practice examples and patterns as they relate to gynecomastia grade and seriousness. The ongoing writing was questioned using the PubMed and MEDLINE data sets — given predefined boundaries and individual audits, 17 investigations were eventually included. Key information focuses included gynecomastia grade, careful intercession, the pace of confusion, including hematoma, seroma, disease, corruption, and channel use. Two-example t-test was used for additional investigation. A sum of 1112 patients went through careful treatment for gynecomastia. Skin-saving mastectomy regardless of liposuction was the most often utilized technique followed by mastectomy with skin decrease. Significant complexity rates went from 0% to 33%, with hematoma development being generally normal (5.8%) followed by seroma (2.4%). There was a higher pace of hematoma/seroma arrangement among creators who regularly used channel situation (9.78% versus 8.36%; P = 0.0051); nonetheless, this is logically inferable from the enormous error in the level of grade III patients found in each gathering (50.23% versus 4.36%; P = 0.0000). As a wide assortment of careful procedures exists for the treatment of gynecomastia, an individualized methodology given gynecomastia grade and patient inclination might help the specialist in giving ideal results. This senior creator's favoured technique for the treatment of gynecomastia is shown in the included calculation. 

[STEPS]
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p><span class="TextRun SCX24498567" lang="EN-US" style="font-size: 14pt; font-family: Times New Roman,serif; line-height: 23px;" xml:lang="EN-US"><span class="NormalTextRun SCX24498567" style="background-color: inherit;">This protocol is designed to allow you to clean large</span></span><span class="TextRun SCX24498567" lang="EN-US" style="font-size: 14pt; font-family: Times New Roman,serif; line-height: 23px;" xml:lang="EN-US"><span class="NormalTextRun SCX24498567" style="background-color: inherit;">r GEOTRACES bottles</span></span><span class="TextRun SCX24498567" lang="EN-US" style="font-size: 14pt; font-family: Times New Roman,serif; line-height: 23px;" xml:lang="EN-US"><span class="NormalTextRun SCX24498567" style="background-color: inherit;"> to the highest standards of cleanliness, but in a time-efficient manner.</span></span></p>

[STEPS]
?. 
?. 
?. 
?. 
?. 
?.
TITLE: Luciferase Activity Assay
AUTHORS: Electra Brunialti, Alessandro Maria Villa, Paolo Ciana

[DESCRIPTION]
Luciferase activity assay on cell extracts using Veritas (Turner) luminometer.

[STEPS]
1. Lysate the cell using 1X Luciferase Cell Culture LysisReagent (Cat. E1531, Promega) following manufacturer instruction; 
3. put 20 µl/well of cell lysate into the white plate (Cat 3912, Corning);
2. prepare Reaction Buffer (100µl for each sample): 500 µM luciferin (Cat E160E, Promega), 20 mM Tricine, 0.1 mM EDTA, 1.07 mM (MgCO3)4·Mg(OH)2, 2.67 mM MgSO4 in H2O, pH 7.8, with 33.3 mM DTT and 530 mM ATP. 
4. turn on the luminometer (Veritas, Turner);
5. let the samples' plate and Reaction Buffer equilibrate at room temperature; Insert the plate into the instrument and prime the tube with the Reaction Buffer;
6. run the protocol on the luminometer (injection volume 100µl, delay before measurement 0s, integration time 10s)
TITLE: LRRK2 PhosphoSens Assay
AUTHORS: Nicolai D. Raig, Stefan Knapp

[DESCRIPTION]
With this enzymatic activity assay we were able to determine IC50 values of published inhibitors aswell as newly synthesized compounds that inhibit LRRK2. The assay is based on the in vitro phosphorylation reaction between the enzymatic active LRRK2(RCKW)-construct and the SOX-based substrate peptide (AQT0615 from AssayQuant Technologies).

[STEPS]
2. Pipett a dilution series of eleven concentrations between 15µM and 0.4nM (calculated with an assay volume of 10µL) of the compounds into white 384-well plates (Greiner 781207) as duplicates with an ECHO acoustic dispenser (Labcyte). Pipett a equivalent of DMSO in two wells per compound as 0% (without protein and compound) and 100%(without compound) control.
3. Dilute the pur20ified LRRK2RCKW to 22nM with the reaction buffer (50mM HEPES buffer (pH7.5), 10mM MgCl2, 1% glycerol, 1mM DTT, 0.2mg/mL BSA, 0.01% Tween20, 5µM AQT0615) and add 10µL to each well, except the 0% control wells,  with a multichannel pipette (Eppendorf). Add   10µL of pure reaction buffer to the 0% control wells.
1. Make 10mM, 100µM and 1.6µM stock solutions of the compounds in DMSO.
4. Pipett 5nL 100mM ATP stock into each well with the ECHO acoustic dispenser (Labcyte).
5. Centrifuge the plates at 1500gn for 2min, before incubating the reaction at room temperature for 4 h.
6. Measure the fluorescence after 360nm excitation at 487nm emission with a PHERAstar plate reader (BMG Labtech).
7. Normalize the response to the average 0% and 100% controls. Calculate the IC50 values via non-linear regression of the log[inhibitor] vs. the normalized response with GraphPad Prism 8.
TITLE: Tissue and blood collection
AUTHORS: michela.deleidi, Bianca Marchetti, Federico Bertoli, Carmela Giachino

[DESCRIPTION]
This protocol details tissue collection from mice.

[STEPS]
SECTION: Tissue and blood collection
1. For immunohistochemistry, mice (5-6 mice/genotype/age-group/treatment) were anesthetized and transcardially perfused with 0.9% NaCl, followed by 4 % paraformaldehyde in phosphate buffer (pH 7.2 at 4°C), the brains and peripheral tissues carefully removed and post-fixed for 2-4 hrs, in 4% paraformaldehyde in phosphate buffer (pH 7.2) and placed in 15% sucrose in the solution of phosphate buffer overnight at 4°C. 

SECTION: Tissue and blood collection
2. Blood was collected into EDTA-treated tubes and serum was separated by centrifugation and stored at -20C.

SECTION: Tissue and blood collection
3. For gene expression and protein analysis, animals were sacrificed by cervical dislocation, the brains and peripheral tissues were quickly isolated and then stored at -80°C until assayed, the tails collected from each mouse were genotyped through PCR followed by electrophoretic analysis, as described in the manufacturer’s instructions (Jackson Laboratory).
TITLE:  Measuring tension, pellet transit, and calcium imaging within cell subtypes in response to direct electrical field stimulation of colon
AUTHORS: Dante  Heredia, Thomas  Gould

[DESCRIPTION]
Measurement of the effects of direct electrical stimulation of the mouse colon

[BEFORE_START]
Make sure there is enough gas.

[GUIDELINES]
Nothing more than in steps.

[STEPS]
1. A ventral midline incision is made and the entire colon is excised and placed into a sylguard-lined dissection dish filled with oxygenated Krebs-Ringers solution.  The colon is then gently flushed of its contents. For all experiments, the colon is perfused with 35-degree, oxygenated Krebs-Ringers for the duration of the expt.
2. For tension, the colon is then drawn over a 1.5-mm diameter fire-polished capillary tube, whose length exceeds that of the colon.  An artificial pellet is mounted to the capillary glass and the colon is positioned with the pellet in the middle. The capillary glass is then fixed to the bottom of the organ bath by the ends protruding from each colonic opening. Suture silk is used to connect three force transducers (model TST125C; Biopac Systems, Santa Barbara, CA) to the proximal, transverse and distal segments of the colon. Resting tension is initially set at 8 mN and monitored using an MP100 interface and recorded on a PC running Acqknowledge software 3.2.6 (Biopac Systems). Two platinum wire electodes are placed on top of proximal or distal colon.  Square wave electrical stimulation, pulsewidth 0.3 ms,  is applied at various frequencies and durations.
3. For pellet transit, the colon is pinned loosely into a dish and placed under a camera.  A mouse fecal pellet that has been dried, then coated with epoxy, and dried again, is placed with forceps into bottom of proximal colon (end of chevrons).The time it takes to begin movement, as well as the speed of movement, through the colon is recorded, both spontaneously and in response to electrical stimulation at various locations and frequencies/durations.
4. For calcium imaging, either Kit-GCaMP6f, ChAT-GCaMP6f, or nNOS-GCaM6f mice are used. In this dissection, the middle colon only is slit from the ventral midline, and pinned with gentle tension.  This is the area of the colon that is imaged. Then calcium responses are measured in response to  electrical stimulation at various locations and frequencies/durations.
TITLE: DNA Extraction for Beetle DNA with Qiagen DNeasy
AUTHORS: Demian F Gomez, Caroline Storer

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">The purpose of this protocol is to extract DNA from beetles using the DNeasy protocol with additional notes to the manufacturer guidelines.</div><div class = "text-block"><span>This protocol is part of the Bark Beetle Mycobiome (BBM) Research Coordination Network. For more information on the BBM international network: Hulcr J, Barnes I, De Beer ZW, Duong TA, Gazis R, Johnson AJ, Jusino MA, Kasson MT, Li Y, Lynch S, Mayers C, Musvuugwa T, Roets F, Seltmann KC, Six D, Vanderpool D, & Villari C. 2020. Bark beetle mycobiome: collaboratively defined research priorities on a widespread insect-fungus symbiosis. Symbiosis 81: 101–113 </span><a href="https://doi.org/10.1007/s13199-020-00686-9" style = "text-decoration:underline;color:blue;cursor:pointer;"><span style = ":;">https://doi.org/10.1007/s13199-020-00686-9</span></a><span>.</span></div></div>

[STEPS]
TITLE: Frequently sampled Insulin glucose tolerance test
AUTHORS: Timothy Nichols, David Clemmons

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Summary: </span></div><div class = "text-block">This assay is used by the DiaComp to measure glucose tolerance and insulin sensitivity in pigs.</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/bidx4kcn.jpg" /></div><div style = "text-align :; float : ;"><img style = "" src = "https://s3.amazonaws.com/pr-journal/bidy4kcn.jpg" /></div><div style = "text-align :; float : ;"><img style = "" src = "https://s3.amazonaws.com/pr-journal/bidz4kcn.jpg" /></div><div style = "text-align :; float : ;"><img style = "" src = "https://s3.amazonaws.com/pr-journal/bid24kcn.jpg" /></div><div style = "text-align :; float : ;"><img style = "" src = "https://s3.amazonaws.com/pr-journal/bid34kcn.jpg" /></div><div class = "text-block"><span style = "font-weight:bold;">Reference:</span></div><div class = "text-block"><span style = "font-style:italic;">1. Bergman RN, Finegood DT, Ader M: Assessment of insulin sensitivity in vivo. Endocr Rev 1985, 6:45-86.</span></div></div>

[STEPS]
?. FSIGT or Bergman analysis Pigs are studied after an overnight fast. The food intake of the animals is monitored for 3 days prior to the fast to ensure adequate carbohydrate intake. Two intravenous catheters are placed, one for sampling and one for infusing glucose and insulin. A bolus of glucose (0.3 gm/kg) is administered as a 50% solution over ~5 min. Blood samples are obtained at –15, -10, -5, -1, 0, 2, 3, 4, 5, 6, 8, 10, 12, 14, 16, and 19 minutes. At 20 minutes an insulin bolus (0.03U/kg) is injected and frequent blood samples for insulin and glucose measurements are collected up to the 180 minute time point. Insulin is measured by RIA (ICN) and glucose is measured on a YSI instrument (Yellow Springs, Ohio). The data were analyzed by the Bergman method to calculate an insulin sensitivity index (SI) using MINMOD Millennium version 6.02.¹
TITLE: Nodule cleaning
AUTHORS: 

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Adapted for high throughput of small samples for sequencing.</div><div class = "text-block">Based on doi:10.1371/journal.pone.0047677</div></div>

[STEPS]
?. Cleanly cut nodules from plant roots and drop into a 2ml Eppendorf tube per plant.
?. Add 1ml 100% EtOH to each tube
?. Incubate for 5 seconds
?. Remove as much liquid as possible without removing loose nodules
?. Add 1ml 6% sodium hypochlorite
?. Remove as much liquid as possible
?. Add 2ml sterile water
?. Remove as much liquid as possible
[Repeat once]
?. Close tube, move to sequencing/culturing
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p>This protocol summarizes the process of primer design for assembly-based cloning methods, such as Gibson or AQUA-cloning. </p>
<p>Well-designed primers are necessary for efficient cloning of any kind.<strong> </strong></p>
<p>This protocol can also be used to design primers for overlap extension PCR of linear fragments. Alternatively, overhangs can also include restriction sites for restriction/ligation based cloning.</p>
<p> </p>

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

[DESCRIPTION]
<p>Now includes media used for phosphate limitation experiments.</p>

[GUIDELINES]
<p>All media should be autoclaved unless otherwise noted</p>
<p> </p>
<p><u>Zobell plates, 100% nutrient (1 L) – for growing host; ~15 mL/plate</u></p>
<p>26 g sea salts</p>
<p>1 g yeast extract</p>
<p>5 g proteose peptone</p>
<p>fill to 1 L qH<sub>2</sub>O</p>
<p>12 g agar</p>
<p> </p>
<p><u>Zobell plates, 20% nutrient (1 L) – for plaque assays; ~10 mL/plate</u></p>
<p>26 g sea salts</p>
<p>0.2 g yeast extract</p>
<p>1 g proteose peptone</p>
<p>fill to 1 L qH<sub>2</sub>O</p>
<p>12 g agar</p>
<p><u> </u></p>
<p><u>PZM (“<em>Psuedoaleromonas</em> Zobell Media”; 50% nutrient Zobell) (1 L) – culture media</u></p>
<p>26 g sea salts</p>
<p>0.5 g yeast extract</p>
<p>2.5 g proteose peptone</p>
<p>fill to 1 L qH<sub>2</sub>O</p>
<p> </p>
<p><u>PZM top agar (1 L) – for plaque assays</u></p>
<p>26 g sea salts</p>
<p>0.5 g yeast extract</p>
<p>2.5 g proteose peptone</p>
<p>fill to 1 L qH<sub>2</sub>O</p>
<p>6 g agarose</p>
<p> </p>
<p><u>MSM (1 L) – phage buffer</u></p>
<p>100 mM NaCl (5.8 g)</p>
<p>81.2 mM MgSO<sub>4</sub>x7H<sub>2</sub>O (20 g)</p>
<p>50 mM Tris base (3.02 g) <strong>OR</strong> 50 mL 1 M Tris (pH 7.6)</p>
<p>fill to 1 L qH<sub>2</sub>O</p>
<p>adjust to pH 7–8 if necessary</p>
<p> </p>
<p><u>Artificial seawater, 26 psu (ASW; 1 L) – dilution buffer</u></p>
<p>26 g sea salts</p>
<p>fill to 1 L qH<sub>2</sub>O</p>
<p> </p>
<p><u>Glucose, 222 mM (1 L)</u></p>
<p>40 g Glucose</p>
<p>fill to 1 L qH<sub>2</sub>O</p>
<p>adjust to pH 7–8 if necessary</p>
<p> </p>
<p><u>Ammonium Sulfate, 166 mM (1 L)</u></p>
<p>22 g (NH<sub>4</sub>)<sub>2</sub>SO<sub>4</sub></p>
<p>fill to 1 L qH<sub>2</sub>O</p>
<p>adjust to pH 7–8 if necessary</p>
<p> </p>
<p><u>Phosphoric Acid, 31 mM (1 L)</u></p>
<p>5.4 g K<sub>2</sub>HPO<sub>4</sub></p>
<p>fill to 1 L qH<sub>2</sub>O</p>
<p>adjust to pH 7–8 if necessary</p>
<p> </p>
<p><u>1%Z+CNP (+P culture media; 1 L)</u></p>
<p>26 g sea salts <strong>OR</strong> use ASW instead of qH<sub>2</sub>O</p>
<p>20 mL PZM</p>
<p>50 mL ammonium sulfate (166 mM; final 8.3 mM)</p>
<p>5 mL phosphoric acid (31 mM; final 0.15 mM)</p>
<p>fill to 1 L qH<sub>2</sub>O <strong>OR</strong> ASW</p>
<p>adjust to pH 7–8 if necessary</p>
<p>autoclave, cool to 60 <strong>°</strong>C</p>
<p>50 mL glucose (222 mM; final 11 mM)</p>
<p> </p>
<p><u>1%Z+CN (-P culture media; 1 L)</u></p>
<p>26 g sea salts <strong>OR</strong> use ASW instead of qH<sub>2</sub>O</p>
<p>20 mL PZM</p>
<p>50 mL ammonium sulfate (166 mM; final 8.3 mM)</p>
<p>fill to 1 L qH<sub>2</sub>O <strong>OR</strong> ASW</p>
<p>adjust to pH 7–8 if necessary</p>
<p>autoclave, cool to 60 <strong>°</strong>C</p>
<p>50 mL glucose (222 mM; final 11 mM)</p>

[STEPS]
?.
TITLE: Passage cells
AUTHORS: Adita Ayu Permanasari

[DESCRIPTION]
Passage Cells

[STEPS]
1. Observe cells under microscope
2. Wipe outside surface of cell culture dish with ethanol cotton and bring it inside Biosafety Cabinet (BSC)
3. Bring medium, PBS, Trypsin-EDTA 2x to BSC
4. Bring autopipetter, disposable pipet (10ml, 2ml), one 15ml tube to BSC
5. Discard old medium
6. Rinse the bottom of dish with 10ml PBS
7. Add 1ml Trypsin-EDTA 2x to dish
8. Incubate the dish in CO2 Incubator 37oC 4min
9. Check under microscope (cell shape become rounded)
10. Neutralize trypsin-EDTA 2x by add 8ml medium
11. Suspend detached cells and transfer to 15ml tube
12. Centrifuge 1200 rpm, 4 min
13. Bring to BSC, discard medium
14. Add 20 ml DMEM and divided each 10 ml to dish (total 2 dish with @50% cells)
TITLE: Genomic DNA extraction from the diatom Pseudo-nitzschia multistriata for Illumina sequencing
AUTHORS: Francesco  Manfellotto, Monia Teresa Russo, Pina Marotta, Rossella Annunziata, Anna Santin, Antonella Ruggiero, Mariella Ferrante

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><span>Genomic DNA extraction from the diatom </span><span style = "font-style:italic;">Pseudo-nitzschia multistriata </span><span>for Illumina sequencing</span></div></div>

[STEPS]
?. Detect the presence/absence of bacteria and collect the cultures as described in: dx.doi.org/10.17504/protocols.io.btt5nnq6
?. Resuspend cells with 500 μL of TE buffer (10 mM TrisHCl pH 7.6 and 1 mM EDTA pH 8.0)
?. Add: 400 mg of 0.2-0.3 mm diameter zirconia/silica beads 500 μL phenol (pH 7.8).
?. Mix with vortex 3 times at 30 Hz for 85 seconds (Each time put sample in ice for 60 seconds before vortex)
?. Centrifuge at 11000 g  for 5 minutes at 4°C.
?. Recover aqueous phase in new 1.5 mL Eppendorf tubes (about 600 μl)
?. Add 500 μL of P.C.I (Phenol:Chloroform:Isoamyl alcohol 25:24:1 v/v) and mix by inversion
?. Centrifuge at 11000 g for 5 minutes at 4°C
?. Move the aqueous phase in a new Eppendorf tube and add 5 µL of RNase-A 10 mg/mL
?. Incubate at 37 °C for 30 minutes.
?. Add 500 μl di P.C.I (Phenol:Chloroform:Isoamyl alcohol 25:24:1 v/v) and mix by inversion.
?. Centrifuge at 11000 g for 5 minutes at 4°C.
?. Move the aqueous phase in a new 2 mL Eppendorf tube and add:50 μL of 3 M NaAc (pH ± 5) 1 mL of ethanol 96% (- 20 °C) 2 μL glycogen (- 20 °C)
?. Incubate over night at -20°C.
?. Centrifuge the samples at 13000 g for 30 minutes at 4°C
?. Discard aqueous phase
?. Add 1 mL ethanol 70% and mix gently by inversion
?. Centrifuge at 13000 g for 10 minutes at 4°C
?. Discard aqueous phase
?. Add 1 mL ethanol 70% and mix gently by inversion
?. Centrifuge at 13000 g for 10 minutes at 4°C
?. Remove aqueous phase and dry pellet at R.T. for at least 20 minutes
?. Add 50 μL of preheated (55°C) TE 1X (pH 8) or sterile MilliQ water
?. Incubate at 55 °C for 20 minutes
?. Quantify DNA concentration by Nanodrop or Qubit
?. Check DNA integrity.Run a small amount of DNA with 1% agarose gel
?. Store DNA at +4 °C, or -20°C for longer storage times
?. Grow cells as described in:  dx.doi.org/10.17504/protocols.io.bgudjws6