mirror of
https://github.com/scinote-eln/scinote-web.git
synced 2024-09-21 23:46:21 +08:00
901 lines
47 KiB
JSON
901 lines
47 KiB
JSON
{
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"experiment": {
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"id": 423,
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"name": "Antibody Purification",
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"description": "Antibody purification involves selective enrichment or specific isolation of antibodies from serum, ascites fluid or cell culture supernatant of a hybridoma cell line. Purification methods range from very crude to highly specific.\r\nOnce they are purified (and possibly after labelling them with an enzyme or fluorescent tag), these antibodies can be used directly to probe the specific antigen in Western blotting, ELISA and other applications.",
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"project_id": 223,
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"created_at": "2018-11-08T10:28:48.389Z",
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"uuid": "37198411-2ad0-4198-b680-47f3b455ce0d"
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},
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{
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{
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"id": 6024,
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},
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"steps": [
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{
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"step": {
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"id": 4287,
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"name": "Before applying a new sample",
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"description": "<!DOCTYPE html PUBLIC \"-//W3C//DTD HTML 4.0 Transitional//EN\" \"http://www.w3.org/TR/REC-html40/loose.dtd\">\n<html><body><p>Re-equilibrate column with buffer until the baseline of the monitored signal is stable. Proteins can be monitored at<strong> 280 nm.</strong></p></body></html>",
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"position": 2,
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"description": "<!DOCTYPE html PUBLIC \"-//W3C//DTD HTML 4.0 Transitional//EN\" \"http://www.w3.org/TR/REC-html40/loose.dtd\">\n<html><body><p><strong>Equilibrate with 2 column volumes of the buffer</strong> at a recommended flow rate (See the table). Apply a sample volume equivalent to between <strong>0.3%</strong> and <strong>4%</strong> of the bed volume. For most applications, the sample volume should not exceed <strong>2%</strong> to achieve high resolution. Smaller sample volumes generally lead to an improved resolution. <br><strong>Elute with 1 column volume of buffer solution</strong> (<strong>10</strong> to <strong>50 mM</strong> sodium phosphate, <strong>150 mM</strong> sodium chloride, <strong>pH 7.0</strong> to <strong>7.4</strong>, or select the buffer in which the sample should be stored or solubilized for the next step). The sample should be fully dissolved. Centrifuge or filter to remove particulate matter.</p></body></html>",
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"name": "Separation options",
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"contents": "eyJkYXRhIjpbWyJGcmFjdGlvbmF0aW9uIHJhbmdlLCBNciAoZ2xvYnVsYXIg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\nIDUwICIsIjIiLCIwLDA3NSJdXX0=\n",
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}
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]
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},
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{
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"step": {
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"id": 5224,
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"name": "For the first time use",
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"description": "<!DOCTYPE html PUBLIC \"-//W3C//DTD HTML 4.0 Transitional//EN\" \"http://www.w3.org/TR/REC-html40/loose.dtd\">\n<html><body>\n<div style=\"text-align: justify;\">Remove storage solution. Applying <strong>2 column</strong> volume of distilled water at room temperature. Use a low flow rate to avoid gap formation. Perform a column performance test (see below) and <strong>determine the limit for maximum pressure over the packed bed is recommended.</strong> If this has already been done, go to <strong>Step 2.</strong><br><br><strong>Column efficiency test (the efficiency and peak asymmetry can be determined as follows): </strong><br><br>\n</div>\n<ol>\n<li style=\"text-align: justify;\">\n<div>Equilibrate the packed column in distilled water or buffer at the flow rate recommended in the product instructions. </div>\n</li>\n<li style=\"text-align: justify;\">\n<div>Inject <strong>2%</strong> acetone (<strong>20mg/mL</strong> in water) in a volume equivalent to <strong>0.2%</strong> to <strong>0.4%</strong> of the geometrical bed volume. </div>\n</li>\n<li style=\"text-align: justify;\">\n<div>Monitor UV absorbance at 280 nm and determine the elution volume for acetone. </div>\n</li>\n<li style=\"text-align: justify;\">\n<div>\n<strong>Calculate column efficiency</strong>, that is, the number of theoretical plates per meter (N/m). <strong>N/m= 5.54 x (<span style=\"color: inherit; font-size: inherit;\">Ve x </span>W<sub>1/2</sub>)<sup>2</sup> x 1/L</strong> ; where V<sub>e</sub> =peak elution (retention) volume; W<sub>1/2</sub>=peak width at half peak height; L= bed height (m)</div>\n</li>\n<li>\n<div style=\"text-align: justify;\">\n<strong>Calculate the asymmetry factor</strong> A<sub>s</sub>= b/a; where a=first half width at <strong>10%</strong> peak height and b=second half width at <strong>10%</strong> peak height</div>\n</li>\n</ol>\n<div> </div>\n</body></html>",
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"position": 0,
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"user_id": 202,
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"created_at": "2018-12-19T07:24:14.116Z",
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"updated_at": "2019-02-20T07:40:29.217Z",
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},
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{
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"id": 3092,
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}
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],
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"assets": [
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{
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"created_at": "2019-02-20T07:39:44.265Z",
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},
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"asset_blob": {
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"filename": "Column_efficiency.jpg"
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}
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}
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],
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}
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],
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"results": []
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},
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{
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"my_module": {
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"id": 2096,
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"name": "Capture",
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"x": 36,
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},
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"outputs": [
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{
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"id": 6023,
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"output_id": 2096
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}
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],
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"created_at": "2018-11-08T10:28:49.025Z",
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"published_on": null,
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"nr_of_linked_children": 0
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},
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"protocol_protocol_keywords": [],
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"steps": [
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{
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"step": {
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"id": 4286,
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"name": "Preparation",
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"description": "<!DOCTYPE html PUBLIC \"-//W3C//DTD HTML 4.0 Transitional//EN\" \"http://www.w3.org/TR/REC-html40/loose.dtd\">\n<html><body><p>Follow the preparation guidelines for capture task.</p></body></html>",
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"position": 0,
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"completed": false,
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"created_at": "2018-11-08T10:28:49.261Z",
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"updated_at": "2018-12-24T09:19:05.983Z",
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"last_modified_by_id": 202,
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},
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"checklists": [
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{
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"checklist": {
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"id": 968,
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"name": "Guideline:",
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"created_at": "2018-12-21T13:27:09.411Z",
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},
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"checklist_items": [
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{
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"id": 3984,
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"text": "Equilibrate all materials to the temperature at which the separation will be performed.",
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||
"checked": false,
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"checklist_id": 968,
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"created_at": "2018-12-21T13:27:09.413Z",
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"position": 0
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},
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{
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||
"id": 3985,
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"text": "Eliminate air by flushing column end pieces with buffer. Ensure no air is trapped under the column net. Close column outlet leaving 1–2 cm of the buffer in the column.",
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||
"checked": false,
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"checklist_id": 968,
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"created_at": "2018-12-21T13:27:09.421Z",
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"position": 1
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},
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{
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"id": 3986,
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||
"text": "Gently resuspend the medium. For media not supplied in suspension, use a medium: buffer ratio of approximately 1:2 to produce a suspension for mixing during rehydration. Avoid using magnetic stirrers since they may damage the matrix.",
|
||
"checked": false,
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||
"checklist_id": 968,
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"created_at": "2018-12-21T13:27:09.427Z",
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"position": 2
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}
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]
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],
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},
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{
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"step": {
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"id": 4279,
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"name": "Stop the pump and close the column outlet.",
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"description": "<!DOCTYPE html PUBLIC \"-//W3C//DTD HTML 4.0 Transitional//EN\" \"http://www.w3.org/TR/REC-html40/loose.dtd\">\n<html><body><p style=\"text-align: justify;\">Remove the top piece and carefully fill the rest of the column with buffer to form an upward meniscus at the top. Insert the adaptor into the column at an angle, ensuring that no air is trapped under the net. Slide the adaptor slowly down the column (the outlet of the adaptor should be open) until the mark is reached. Lock the adaptor in position. Connect the column to the pump and begin equilibration. Re-position the adaptor if necessary.</p></body></html>",
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"position": 2,
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"completed": false,
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"created_at": "2018-11-08T10:28:49.069Z",
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"updated_at": "2018-12-21T13:29:08.411Z",
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},
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},
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{
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"step": {
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||
"id": 4283,
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||
"name": "Column fill and setting the column outlet.",
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||
"description": "<!DOCTYPE html PUBLIC \"-//W3C//DTD HTML 4.0 Transitional//EN\" \"http://www.w3.org/TR/REC-html40/loose.dtd\">\n<html><body><p style=\"text-align: justify;\">Estimate the amount of resuspended medium required and our the required volume of the medium into the column. Pouring down a glass rod held against the wall of the column will minimize the introduction of air bubbles. <strong>Immediately fill the column with buffer</strong>. Mount the column top piece and connect to a pump. Open the column outlet and set the pump to the desired flow rate. If the recommended flow rate cannot be obtained, use the maximum flow rate the pump can deliver. <strong>Do not exceed the maximum operating pressure of the medium or column.</strong> Maintain the packing flow rate for at least <strong>3 column volumes</strong> after a constant bed height is obtained. Mark the bed height on the column. Do not exceed <strong>75%</strong> of the packing flow rate during any purification.</p></body></html>",
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"position": 1,
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"completed": false,
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},
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{
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||
"checked": false,
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||
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||
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"position": 2
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},
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{
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||
"id": 3990,
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"text": "Add 1.5 mL of Bradford reagent to each tube and mix well.",
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"checked": false,
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||
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||
"position": 3
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||
},
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||
{
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"id": 3991,
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||
"text": "Incubate at room temperature for at least 5 min.",
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||
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||
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||
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|
||
"position": 4
|
||
},
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||
{
|
||
"id": 3992,
|
||
"text": "Absorbance will increase over time; samples should incubate at RT for no more than 1 h.",
|
||
"checked": false,
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||
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||
"created_at": "2018-12-21T13:35:09.703Z",
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||
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||
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|
||
"position": 5
|
||
},
|
||
{
|
||
"id": 3993,
|
||
"text": "Measure absorbance at 595 nm.",
|
||
"checked": false,
|
||
"checklist_id": 969,
|
||
"created_at": "2018-12-21T13:35:09.710Z",
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||
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||
"created_by_id": null,
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||
"last_modified_by_id": null,
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||
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|
||
}
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]
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}
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],
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||
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}
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],
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"my_module_groups": [
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{
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||
"id": 1190,
|
||
"created_at": "2018-12-21T13:25:20.378Z",
|
||
"updated_at": "2018-12-21T13:25:20.378Z",
|
||
"created_by_id": 202,
|
||
"experiment_id": 423
|
||
}
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