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

[DESCRIPTION]
This protocol is a group of the following 5 methods from the NEB website:<br />1.&nbsp;<a href="https://www.neb.com/protocols/2012/11/15/nebnext-end-prep-e7370" target="_blank">NEBNext End Prep </a> <br />2. <a href="https://www.neb.com/protocols/2012/11/15/adaptor-ligation-e7370" target="_blank">Adaptor Ligation</a><br />3.&nbsp;<a href="https://www.neb.com/protocols/2012/11/15/size-selection-of-adaptor-ligated-dna-e7370" target="_blank">Size Selection of Adaptor-ligated DNA</a><br />4.&nbsp;<a href="https://www.neb.com/protocols/2012/11/19/pcr-amplification-e7370" target="_blank">PCR Amplification</a><br />5.&nbsp;<a href="https://www.neb.com/protocols/2012/11/19/cleanup-of-pcr-amplification-e7370" target="_blank">Cleanup of PCR Amplification</a>

[GUIDELINES]
<p>The NEBNext Ultra DNA Library Prep Kit for Illumina contains enzymes and buffers that are ideal to convert a small amount of DNA input into indexed libraries for next-generation sequencing on the Illumina platform (Illumina, Inc).  <br /> <br /><strong>Adaptor Ligation:</strong><br /><br />If DNA input is &lt; 100 ng, dilute the NEBNext Adaptor for Illumina* 1:10 and use immediately to a final concentration of 1.5 μM.<br /><br /><strong>Size Selection:<br /></strong><br />This protocol is suitable for input amounts of 50ng or higher. For lower input amounts we recommend that the protocol without size selection be followed.<br /><br />This size selection is for libraries with 200 bp inserts only and is based on a starting volume of 100 µl. For libraries with different size fragment inserts, refer to Table 1 for  the appropriate volume of beads to be added. <br /><br />Table 1: Recommended conditions for bead based size selection.<strong> </strong></p>
<table>
<tbody>
<tr>
<td rowspan="2" valign="middle"><strong>LIBRARY</strong><br /><strong>PARAMETERS</strong></td>
<td valign="middle"><strong>APPROXIMATE</strong><br /><strong>INSERT SIZE</strong></td>
<td><strong>150 bp</strong></td>
<td><strong>200 bp</strong></td>
<td><strong>250 bp</strong></td>
<td><strong>300-400 bp</strong></td>
<td><strong>400-500 bp</strong></td>
<td><strong>500-700 bp</strong></td>
</tr>
<tr>
<td valign="middle">Total Library Size (insert + adaptor)</td>
<td>270 bp</td>
<td>320 bp</td>
<td>400 bp</td>
<td>400-500 bp</td>
<td>500-600 bp</td>
<td>600-800 bp</td>
</tr>
<tr>
<td rowspan="2" valign="middle"><strong>VOLUME TO</strong><br /><strong>BE ADDED (μl)</strong></td>
<td>1st Bead Selection</td>
<td>65</td>
<td>55</td>
<td>45</td>
<td>40</td>
<td>35</td>
<td>30</td>
</tr>
<tr>
<td>2nd Bead Selection</td>
<td>25</td>
<td>25</td>
<td>25</td>
<td>20</td>
<td>15</td>
<td>15</td>
</tr>
</tbody>
</table>
<p><br /><strong><br /></strong>Figure 1: Examples of libraries prepared with human IMR-90 gDNA.<strong><br /><img src="http://i.imgur.com/Jg1AiWp.png" alt="" width="700" height="493" /><br /><img src="http://i.imgur.com/Ytnm7tX.png" alt="" width="700" height="512" /><br /></strong></p>

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

[DESCRIPTION]
<p>We have available several plasmids designed for expression of Cas9, guide RNA, chloramphenicol acetyl-transferase, GFP, and beta-lactamase in Micromonas CCMP1545. For protein expression we used the promotor and 3' end elements from the endogenous RPS9 gene, and codon optimized the coding region.  For expression of guide RNAs we used the Micromonas U6 snRNA promoter.  DNA is available by contacting M. Ares &lt;ares@ucsc.edu&gt;.</p>
<div class="">We have constructed and sequence verified 9 plasmids which we would like to make available to others attempting to detect transformation of DNA into Micromonas. Using the CCMP1545 genome as a source for the U6 promoter sequence, and for the promoter and 3' UTR sequences of ribosomal protein RPS9, we built the following plasmids:</div>
<div class=""> </div>
<div class=""><strong>CRISPR/Cas9 plasmids for Micromonas</strong></div>
<div class="">1. Mp U6 promoter driving Bae cassette for guide RNA expression in pUC13</div>
<div class="">2. Mp RPS9-Cas9SV40-RPS9 in pUC13</div>
<div class="">3. Both Mp U6 promoter driving Bae cassette for guide RNA expression and Mp RPS9-Cas9SV40-RPS9 in pUC13</div>
<div class=""> </div>
<div class="">These first three plasmids were anticipated to enable stable incorporation of transgenes at specific genomic locations. BaeI is a type IIS restriction enzyme that leaves noncompatible sticky ends. In the context of the gRNA cassette, a pair of 24 nt oligos designed to have the sticky ends compatible with BaeI cleaved plasmid are annealed and cloned into the plasmid, replacing the BaeI cassette while adding the 20 nt target complementary sequence of the desired guide RNA.</div>
<div class=""> </div>
<div class=""><strong>Selectable/Detectable Marker genes</strong></div>
<div class="">4. Mp RPS9-codon optimized GFPsv40-RPS9 in pUC13</div>
<div class="">5. Mp RPS9-codon optimized chloramphenicol acetyltransferase-RPS9 in pUC13<br class="" />
<div>6. Mp RPS9-codon optimized beta-lactamase-RPS9 in pUC13</div>
<div> </div>
<div>These three plasmids have the indicated coding regions codon optimized for Micromonas flanked by RPS9 promoter and 3' end sequences. In the case of GFP, a nuclear localization signal from SV40 has been added to the C-terminus.</div>
<div> </div>
<div><strong>Plasmids for Agrobacterium-mediated gene transfer</strong></div>
<div>
<div class="">7. Mp RPS9-codon optimized GFPsv40-RPS9 in pOSCAR</div>
<div class="">8. Mp RPS9-codon optimized chloramphenicol acetyltransferase-RPS9 in pOSCAR</div>
<div class="">9. Mp RPS9-codon optimized beta-lactamase-RPS9 in pOSCAR</div>
</div>
</div>

[STEPS]
TITLE: No Title
AUTHORS: 

[BEFORE_START]
<div class="page" title="Page 2">
<div class="section">
<div class="layoutArea">
<div class="column">
<p>Make sure that the desired protein has the correct restriction sites and is in frame with the intein in the purification vector.</p>
<ol>
<li>
<p>If that is not the case natively, design primers using the sequences in the following table.</p>
</li>
<li><img id="s-mce-img" class="s-mce-img" src="https://s3.amazonaws.com/pr-journal/gs2e7qw.png" width="566" height="347" data-src="https://s3.amazonaws.com/pr-journal/gs2e7qw.png" data-ofn="Screen Shot 2016-11-25 at 13.21.46.png" /></li>
<li>Then perform a Phusion PCR with the designed primers</li>
</ol>
</div>
</div>
</div>
</div>

[GUIDELINES]
<p><span style="text-decoration: underline;"><strong>IMPACT Column Buffer (1L)</strong></span></p>
<p> </p>
<table style="height: 162px;" width="342">
<tbody>
<tr>
<td style="width: 163px;"><strong>Reagent</strong></td>
<td style="width: 163px;"><strong>Volume</strong></td>
</tr>
<tr>
<td style="width: 163px;">Tris-HCl 7-8 pH 1M</td>
<td style="width: 163px;">20 ml</td>
</tr>
<tr>
<td style="width: 163px;">NaCl 5M</td>
<td style="width: 163px;">100 ml</td>
</tr>
<tr>
<td style="width: 163px;">EDTA 0.5M</td>
<td style="width: 163px;">2 ml</td>
</tr>
<tr>
<td style="width: 163px;">Tween-20</td>
<td style="width: 163px;">2 ml</td>
</tr>
<tr>
<td style="width: 163px;">H<sub>2</sub>O</td>
<td style="width: 163px;">-&gt; 1000ml</td>
</tr>
</tbody>
</table>
<p> </p>
<p><span style="text-decoration: underline;"><strong>IMPACT Cleavage Buffer (100ml)</strong></span></p>
<p> </p>
<table style="height: 162px;" width="342">
<tbody>
<tr>
<td style="width: 163px;"><strong>Reagent</strong></td>
<td style="width: 163px;"><strong>Volume</strong></td>
</tr>
<tr>
<td style="width: 163px;">Tris-HCl 7-8 pH 1M</td>
<td style="width: 163px;">2 ml</td>
</tr>
<tr>
<td style="width: 163px;">NaCl 5M</td>
<td style="width: 163px;">10 ml</td>
</tr>
<tr>
<td style="width: 163px;">EDTA 0.5M</td>
<td style="width: 163px;">0.2 ml</td>
</tr>
<tr>
<td style="width: 163px;">DTT</td>
<td style="width: 163px;">0.2 ml</td>
</tr>
<tr>
<td style="width: 163px;">H<sub>2</sub>O</td>
<td style="width: 163px;">-&gt; 100ml<br /><br /></td>
</tr>
</tbody>
</table>

[STEPS]
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?.
TITLE: Bueno Sopaipillas 
AUTHORS: Chasz Griego

[DESCRIPTION]
Sopaipillas are delicious, oily dough pockets. A New Mexican staple. Eat them as a dessert with honey, or as an entree, stuffing the sopaipilla with meat, beans, cheese, rice, etc. The options are near limitless! This protocol describes the steps to make sopaipillas using a box of mix from the BUENO foods company.

[STEPS]
SECTION: Ingredients
1. YOU WILL NEED
1 Box of Bueno Sopaipilla Mix
1 Tbsp vegetable oil 
3/4 cup warm water
3 cups vegetable oil (for frying)

SECTION: Make Dough
2. EMPTY entire contents of sopaipilla mix into a large bowl.
SECTION: Make Dough
3. ADD 1 Tbsp. oil. Gradually add water.
 

SECTION: Make Dough
4. KNEAD the dough on a lightly floured surface for 5 min until smooth.

SECTION: Make Dough
5. STORE the dough in a plastic bag, making sure the bag is wrapped tightly around the dough. Let sit for 25 min in a dry, warm place.

Room temperature   

SECTION: Make Dough
6. ROLL dough into a square   thick.

SECTION: Make Dough
7. CUT into squares approximately   by   .

SECTION: Heat
8. HEAT 3 cups of vegetable oil to 350 °F - 400 °F 

 

SECTION: Fry
9. PLACE squares of dough, one at a time, in oil.

SECTION: Fry
10. FRY until the sopaipilla puffs on one side. Turn and fry until golden brown.

SECTION: Fry
11. DRAIN on paper towel. 
 

SECTION: Fry
12. SERVE with honey, powdered sugar, or cinnamon sugar. Or, make stuffed sopaipillas.
TITLE: Transgenic mouse colony maintenance
AUTHORS: Ariadna Laguna, Iria Carballo-Carbajal, Miquel Vila

[DESCRIPTION]
Methods for the generation and maintenance of transgenic tyrosinase mice

[STEPS]
1. Pronuclear microinjection of the human
tyrosinase complementary DNA fused to the rat tyrosine hydroxylase promoter into C57Bl6-SJL (RRID:MGI:5656718) mouse zygotes  
2. Selection of positive lines and backcrossing for 8-10 generations using
C57BL/6J mice (Charles River; RRID:MGI:3028467) 
3. Maintainance in heterozygosis by crossing heterozygous males with C57BL76J females
4. Housed two to five per cage with ad libitum
access to food and water during a 12 hours (h) light/dark cycle (light-on 8
a.m.)
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
This protocol is used for those that do not have a&nbsp;&ge;20,000 x g capable centrifuge.

[BEFORE_START]
Add RNase A solution to Buffer P1, mix, and store at 2&ndash;8&deg;C.<br />Optional: Add LyseBlue&reg; reagent to Buffer P1 at a ratio of 1:1000.<br />Prechill Buffer P3 at 4&deg;C. Check Buffer P2 for SDS precipitation.<br /> Isopropanol and 70% ethanol are required.

[STEPS]
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?.
TITLE: The leading role of evidence-based practices in the treatment of patients with substance use disorders: A systematic review
AUTHORS: qasirabbas

[DESCRIPTION]
Objectives: This study aims to explore the effectiveness of motivational interviewing (MI), motivational enhancement therapy (MET), and cognitive behaviour therapy (CBT) for patients with substance use disorders and to estimate the effect of this comparison in the patient care setting.
Methods: In database electronic search results, 314 articles were found, 273 were excluded, 41 were assessed for eligibility, and 16 were reviewed. In this systematic review, eleven databases were searched between 2001 to 2021 were explored from September 2021 to February 2022. Most of the studies were conducted in USA=6 and UK=3, and limited studies were conducted in Germany=1, England=1, Australia=1, Korea=1, South Africa=1, Alabama=1, and Miami=1.
Result: All studies were intervention-based, and six RCTs provided pre-and-post comparisons between experimental and controls. Eight studies were conducted using MI and CBT and produced substantial improvement among patients. Five studies produced effective and significant results with a combination of MET and CBT, such as adherence to treatment, reduced secondary symptoms, and MET supported the efficacy of CBT. Three RCTs supported MET and CBT effective practices in combination. Two studies combining MI, MET, and CBT reported significant results simultaneously addressing multiple patients’ variables.
Conclusion: All studies were heterogeneous with different interventions, case controls, samples, and outcomes. MI produced short-term treatment outcomes and played a supportive and helping role in sustaining motivation. MET was an effective therapeutic intervention that significantly addressed the inadequate motivations and enhanced motivations for treatment. CBT improved short-term and remained effective in the long term to address cognitive and behavioral setbacks.

[STEPS]
SECTION: The leading role of evidence-based practices in the treatment of patients with substance use disorders: A systematic review
1.
TITLE: An on-site adaptable test for rapid and sensitive detection of Potato mop-top virus, a soil-borne virus of potato (Solanum tuberosum)
AUTHORS: Ying  Zhai, Bryant Davenport, Keith Schuetz,   Hanu R Pappu

[DESCRIPTION]
Potato mop-top virus(PMTV) is considered an emerging threat to potato production in the United States. PMTV is transmitted by a soil-borne protist, Spongospora subterranean. Rapid, accurate, and sensitive detection of PMTV in leaves and tubers is an essential component in PMTV management program. A rapid test that can be adapted to in-field, on-site testing with minimal sample manipulation could help in ensuring the sanitary status of the produce in situations such as certification programs and shipping point inspections. Toward that goal, a rapid and highly sensitive recombinase polymerase amplification (RPA)-based test was developed for PMTV detection in potato tubers. The test combines the convenience of RPA assay with a simple sample extraction procedure, making it amenable to rapid on-site diagnosis of PMTV. Furthermore, the assay was duplexed with a plant internal control to monitor sample extraction and RPA reaction performance. The method described could detect as little as 10 fg of PMTV RNA transcript in various potato tissues, the diagnostic limit of detection (LOQ) similar to that of traditional molecular methods. 

[STEPS]
1. Cut core tubers with blade. Take 3 - 4 cores per tuber.
2. Extract with GEB extraction buffer at a ratio of 1:2 (w:v) in a mesh extraction bag (0.3 g/3 mL). Let rest for 5 minutes at room temperature.
3. Remove one colored PD1 filled tube for each sample being tested. Individual tubes may be cut from the strip of tubes using scissors.
4. Transfer 5 μL of sample extract into the tube containing PD1 diluent and mix well.
5. Press the “Execute Reaction” button on the AmpliFire®. Then Scan PMTV Product Code.
6. Remove a canister of reaction pellets from the white foil pouch labeled with the barcode. Then remove a strip of reaction pellets from the desiccated container. Note: Reaction Pellets are light sensitive. Immediately place remaining reaction pellets back into the desiccated tube and then insert the desiccant tube into the foil pouch to protect from light.
7. Transfer 25 μL from the colored tube from step 4, into the reaction pellet (clear tube). Mix well and spin down.
8. Press “Start” on the AmpliFire. Immediately follow the prompts to add your reactions, press “OK”, and put the lid down.
9. After 4 minutes of incubation remove the reaction(s) from the AmpliFire. Quickly mix, spin, and reinsert the reaction(s) into the AmpliFire to continue monitoring results. Take care to ensure the tubes are in their original positions and orientations.
10. The test lasts 20 minutes and the results will be visible on the screen, and should be interpreted as follows: Blue curve = FAM = PMTV. Red curve = CalRed = Internal control. ( + ) = Positive for PMTV ( - ) = PMTV not detected ( ! ) = Invalid
11. Note: Adapted and modified from the manual of Agdia AmplifyRP® XRT for PMTV Rapid RNA Amplification Test Kit, Product No. XCS 12501
TITLE: Artificial saliva
AUTHORS: Bjorn  Bartholdy, a.g.henry  

[DESCRIPTION]
Creating an artificial saliva solution for oral biofilm growth.

This is a modified version of Sissons et al. 1991.

[BEFORE_START]
Mix the solution under a fumehood. It can smell pretty bad.
Also have the incubator under a fumehood, if possible. I had no such luxury...

[GUIDELINES]
This protocol explains how to mix 1000 mL  of artificial saliva.


[STEPS]
1. Add 300 mL distilled (or deionized) dH2O to a 1000 mL  beaker, with stirring and heat 60 °C .

2. Add:

-  2.5 g   
-  5 g   
- 10 g   
- 5 g   

Let the reagents completely dissolve before continuing to the next step

3. Add:

- 2.5 g   
- 0.35 g   
- 0.2 g   
- 0.74 g   
- 0.54 g   
- 2.5 mg   

5. Adjust to pH 7 with   and stirring

6. Transfer to two 1000 ml bottles, so half of each bottle is filled. 

Autoclave at 121 °C ,   for 15 min minutes
 

7. Once the solution has cooled, add:

1 mg   
0.3 g   
0.17 g   


8. Store in fridge at ca. 4 °C 

Occasionally test the pH to ensure it stays around pH 7 

4. Add the remaining 700 mL  distilled (or deionized) dH2O
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p>For convenience and for PCR products that are challenging to purify with high efficiency (<em>e.g</em>., chemically modified DNAs), it is often desirable to quantitate synthesized DNA directly from a PCR reaction. Here we describe the use of a high-sensitivity Quant-iT™ PicoGreen® dye-based fluorescence assay to quantitate PCR-synthesized, double-stranded, low molecular weight, 5’-modified DNA probes in the presence of single-stranded primers and deoxyribonucleotides.<sup>1</sup></p>

[BEFORE_START]
<ol>
<li>Calibrate your fluorometer as directed by manufacturer’s instructions. For example, if using a Thermo Fisher Scientific Qubit 2.0 fluorometer and Quant-iT™ dsDNA High-Sensitivity (HS) Assay Kit, perform a two-point calibration with samples containing 0 ng and 100 ng λ DNA, provided.</li>
<li>It is helpful to retain an aliquot (2 µL) of a representative PCR reaction before amplification. Store on ice. Such serves as a blank for subsequent PCR quantitation.</li>
</ol>

[GUIDELINES]
<p>Protocol development supported by grants from the NIH (<a href="&quot;https://projectreporter.nih.gov/project_info_description.cfm?a" target="_blank">1R15GM104833-01</a>) and NSF (<a href="https://www.nsf.gov/awardsearch/showAward?AWD_ID=1714778&amp;HistoricalAwards=false" target="_blank">MCB-1714778</a>).</p>

[STEPS]
?. 
?. 
?. 
?. 
?. 
?.
TITLE: Quarantine and heat treatment for Hymenochirus boettgeri infected with Bd
AUTHORS: Tamilie Carvalho, Catherine Si, Rebecca A. Clemons, Evelyn Faust, Timothy Y. James

[DESCRIPTION]
This protocol outlines the procedures for conducting heat treatment for clearing Batrachochytrium dendrobatidis (Bd) infection of H. boettgeri. It ensures that individuals are free of Bd for use in subsequent experiments.

[GUIDELINES]
Conclusion:

This protocol outlines the steps for quarantining and conducting heat treatment on H. boettgeri frogs infected with Bd to eliminate the infection.

[STEPS]
SECTION: Quarantine setup
1. Place Bd-positive frogs in 38 L standard system glass tanks, condition the water, feed the animals, and conduct water changes as described in “Housing and Care for Hymenochirus boettgeri” protocol.
 

SECTION: Heat Treatment
2. Gradually increase the water temperature in the quarantine standard system tanks from room temperature to 30 °C over a period of 10 days using an aquarium heater.

SECTION: Heat Treatment
3. Maintain the water temperature at 30 °C for 14400 min.

SECTION: Heat Treatment
4. Gradually decrease the water temperature from 30 °C to 25 °C over a period of five days.

SECTION: Heat Treatment
5. Monitor and maintain consistent temperature levels throughout the treatment process.
SECTION: Allocation to permanent aquaria
6. Monitor the animals for an additional 10 days after the heat treatment, and then transfer them to permanent aquaria.
TITLE: Case - Measurement of Food Consumption and Body Weight
AUTHORS: Henri Brunengraber

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><span style = "font-weight:bold;">Summary: </span></div><div class = "text-block">During the course of development or during a diet study the amount of food and body weights are measured 1-2 times per week.</div></div>

[STEPS]
?. Mice are marked by ear notches. They are weighed and the weight is recorded.
?. At 9:00AM, mice are placed in a clean cage (2-3 mice/cage because singly housed mice are stressed). The amount of food is weighed and recorded.
?. The next day at 9:00AM the food and mice are weighed and measurements are recorded.
?. To calculate food intake. Subtract day 3 values from day 2 values and divide by the total weight of the mice in the cage.
?. The data is represented as an average over the course of diet study or each week over time.
TITLE: DMSO stock preparation
AUTHORS: Akashdutta  

[DESCRIPTION]
Bacterial cultures in the form of agar plates or liquid cultures are not suitable for long term storage at low temperatures such as 4oC. Long-term storage of bacteria is best at temperatures around -80oC. However, agar plates and liquid cultures cannot be stored at such temperatures, as the water crystallizes in and around the bacterial cells, rupturing and killing them. Glycerol or DMSO (anti-freezing agents) is added to the media to store samples at such temperatures. 


[STEPS]
1. From the streaked plate of the bacteria, pick one colony and inoculate into a 250 mL flask containing 100 mL  culture.

2. Grow the culture till it reaches OD∼ 0.6-0.8
3. Take the 100 mL  culture and transfer 40 mL each into two falcon tubes.

4. Leave the remaining 20 mL culture in the flask. To that, add 80 mL of micronutrient enriched BG-11 media.

5. Take the two falcon tubes and centrifuge them at 5000 rpm, 7 min, 25 °C .

6. Discard the supernatants of the centrifuged pellets.

7. Add 2 mL  of BG-11 into each of the emptied falcon tubes. Use a 1 mL  micropipette to mix such that the pellet is resuspended into a concentrated culture.

8. Add 140 µL of 7% DMSO.

9. Take 4 cryovials. Add 1 mL  of the liquid in each cryovial.

10. Flash-freeze the vials using liquid nitrogen.
11. Store the stocks at  -80 °C
TITLE: DNA extraction (Zymo kit)
AUTHORS: Tsu-Chun  Hung

[DESCRIPTION]
DNA extraction (Zymo kit)

[STEPS]
SECTION: Sample Collection
1. Add 200 µL of 1mm beads to 1.5ml enppendorf tube

SECTION: DNA purification
2. 
SECTION: DNA purification
1. 
SECTION: Sample Collection
1. 
SECTION: Sample Collection
3. Add 600 µL of lysis solution to 1.5ml enppendorf tube
SECTION: Sample Collection
2. Add 200 µL of 0.5mm beads to 1.5ml enppendorf tube

SECTION: Sample Collection
4. Collect 9-10 µg of sample to 1.5ml enppendorf tube
SECTION: Sample crush
5. Put 1.5ml eppendorf tube in the mixmill for sample crush; at condition: 30 rpm/s, for 4mins 4 min 
SECTION: Centrifugation
6. Put 1.5ml eppendorf tube in centrifuge for centrifugation, at this condition:17.0 x g, 3 min, 25 °C 

SECTION: DNA purification
7. Add 600 µL of DNA binding buffer to 1st well of 96 deep well plate 

SECTION: DNA purification
7.2. Add 200 µL of sample (lysate) from 15ml centrifuge microtube to 1st well of 96 deep well plate
 

SECTION: DNA purification
7.1. Add 25 µL of magnetic beads to 1st well of 96 deep well plate
 


SECTION: DNA purification
8. Add 500 µL of Wash 1 to 2nd well of 96 deep well plate
SECTION: DNA purification
9. Add 500 µL of Wash 2 to 3rd well of 96 deep well plate
SECTION: DNA purification
10. Add 500 µL of Wash 2 to 4th well of 96 deep well plate
SECTION: DNA purification
11. Add 100 µL of water to 5th well of 96 deep well plate
SECTION: DNA purification
12. Put in the automated DNA extraction machine
TITLE: Ampicillin 100 mg/mL Stock Solution 
AUTHORS: Bailey  Clark

[DESCRIPTION]
The abstract will be added later.

[STEPS]
SECTION: Stock Solution Preparation
1. Weigh 100 mg  (0.1 g ) of Ampicillin. 

SECTION: Stock Solution Preparation
2. Add the Ampicillin to a 1.5 mL tube.
SECTION: Stock Solution Preparation
3. Add deionized water to the 1 mL level on the tube.
SECTION: Stock Solution Preparation
4. Vortex the tube for 5 s ,  repeat 3 times. 

SECTION: Stock Solution Preparation
5. Store the tube in the -20°C freezer.