{"_id":"57bb90004a92b80e0073d408","parentDoc":null,"category":{"_id":"568f0e73bdb9260d00149d8c","pages":["568f0ea29ebef90d0087276a","568f0eafb700ce0d002f4a32","568f0ebbbdb9260d00149d8d","568f0edeb700ce0d002f4a34","568f0ef3bdb9260d00149d8f","568f12099ebef90d0087276d","568f121abeb2700d00471839","568f20e49ebef90d0087277e","568f446d9ebef90d0087278f","56c79a728bf67e0d00734767","56cb8830c675f50b00a4b834","56d47e99a4a9211b00c8f0d2","56d4af6c1c4de4130005d74e","56d52a131c4de4130005d80b","56de001b3db43f2000d9a765","56de1087f25ce60e002e31ad","56e314c46e602e0e00700b35"],"__v":17,"project":"5476bf0f817e8d080031f988","version":"5476bf10817e8d080031f98b","sync":{"url":"","isSync":false},"reference":false,"createdAt":"2016-01-08T01:18:43.343Z","from_sync":false,"order":1,"slug":"protocols","title":"Premade Protocols"},"user":"56f57b8eab3f610e000a6596","version":{"_id":"5476bf10817e8d080031f98b","__v":17,"project":"5476bf0f817e8d080031f988","createdAt":"2014-11-27T06:05:04.263Z","releaseDate":"2014-11-27T06:05:04.263Z","categories":["5476bf10817e8d080031f98c","5477c46cf3736008009e9eb5","5477c474f3736008009e9eb6","5477c47ef3736008009e9eb7","5477c48ff3736008009e9eb8","5477c4948deb230800808bf0","54e68328154f8e0d0007b55c","54e90194c8e0c00d007ac061","54eed2275bf74a0d00ef4076","54f7a7be0a3cbb0d00d666fb","559b0ebf7ae7f80d0096d871","55d697f9ae529e0d00d34f03","562d4dcc8c6e5a0d00d6ed1d","562e591c4376430d006f17e0","568f0e73bdb9260d00149d8c","5719542aac1e2e0e001834c6","57a14a8ed778850e0047e230"],"is_deprecated":false,"is_hidden":false,"is_beta":false,"is_stable":true,"codename":"","version_clean":"1.0.0","version":"1.0"},"__v":0,"project":"5476bf0f817e8d080031f988","updates":[],"next":{"pages":[],"description":""},"createdAt":"2016-08-22T23:51:28.392Z","link_external":false,"link_url":"","githubsync":"","sync_unique":"","hidden":true,"api":{"results":{"codes":[]},"settings":"","auth":"required","params":[],"url":""},"isReference":false,"order":22,"body":"[block:callout]\n{\n  \"type\": \"warning\",\n  \"title\": \"Bacterial Aliquot Expiration\",\n  \"body\": \"Please note that bacterial aliquots produced using this protocol will expire and be discarded 3 weeks after their creation. Ordering minipreps, maxipreps, or creating glycerol stocks is recommended within this time frame to preserve successfully transformed bacteria.\"\n}\n[/block]\n\n[block:api-header]\n{\n  \"type\": \"basic\",\n  \"title\": \"Description\"\n}\n[/block]\nThe Transform, Spread, Pick protocol is used for transformation of plasmid DNA into Zymo Mix & Go competent cells followed by colony growth, picking, and OD600 validation. It is often followed by the [Sequencing](doc:sequencing)  protocol for sequence verification, and [Plasmid Mini Prep](doc:plasmid-mini-prep) ] or [Plasmid Maxi Prep](doc:plasmid-maxi-prep) protocols to produce aliquots of purified plasmid. \n[block:api-header]\n{\n  \"type\": \"basic\",\n  \"title\": \"Sample requirements\"\n}\n[/block]\n\n[block:parameters]\n{\n  \"data\": {\n    \"h-0\": \"Sample\",\n    \"h-1\": \"Explanation\",\n    \"0-0\": \"DNA to Transform\",\n    \"0-1\": \"An aliquot of plasmid DNA that you wish to transform into chemically competent *E. coli* cells (see below for cell choices).\"\n  },\n  \"cols\": 2,\n  \"rows\": 1\n}\n[/block]\n\n[block:api-header]\n{\n  \"type\": \"basic\",\n  \"title\": \"Required parameters\"\n}\n[/block]\n\n[block:parameters]\n{\n  \"data\": {\n    \"h-0\": \"Parameter\",\n    \"h-1\": \"Explanation\",\n    \"0-0\": \"Bacteria for Transformation\",\n    \"0-1\": \"The bacteria into which you would like to insert your plasmid DNA. All strains are Zymo Mix & Go bacteria designed for quick transformation without the heat shock step. Available strains are Zymo DH5a, Zymo 10B, and Zymo JM109.\",\n    \"1-0\": \"Volume to Transform\",\n    \"2-0\": \"Colonies\",\n    \"1-1\": \"Volume of DNA to transform into competent cells. Must be between 2-5 uL.\",\n    \"2-1\": \"Number of colonies to pick per transformation. Must be 1-10.\",\n    \"3-0\": \"Recover with SOC Medium at 37C\",\n    \"3-1\": \"Adds 400 uL SOC medium to bacteria for a post-transformation recovery at 37C for 50 minutes when selected.\",\n    \"4-0\": \"Group Label\",\n    \"4-1\": \"The label for one transformation using the specified DNA to Transform. Each group will be transformed using the same specified DNA aliquot(s) and use the same antibiotic for agar plate and growth media.\",\n    \"5-0\": \"Antibiotic for Growth Media and Agar Plate\",\n    \"5-1\": \"The antibiotic used on the agar plates for spreading the bacteria after transformation, and in the growth media after colony picking. Choices include 100 ug/mL Ampicilin, 50 ug/mL Kanamycin, 100 ug/mL Spectinomycin, 25 ug/mL Chloramphenicol, or No Antibiotic.\"\n  },\n  \"cols\": 2,\n  \"rows\": 6\n}\n[/block]\n\n[block:api-header]\n{\n  \"type\": \"basic\",\n  \"title\": \"Protocol Outputs\"\n}\n[/block]\nThis protocol outputs new aliquots of bacterial colonies picked into growth media which are stored in your inventory at 4°C. Please note that these bacterial aliquots expire and will be discarded 3 weeks after their creation. Any aliquots of DNA used in the making of these colonies will be returned to the storage condition from which they came.\n\nAn annotated autopick image is returned showing where on the agar plate colonies were picked. OD600 data is returned for the growth plate to verify colony growth after picking.\n[block:api-header]\n{\n  \"type\": \"basic\",\n  \"title\": \"Recommended guidelines\"\n}\n[/block]\nZymo Research recommends keeping the added volume of DNA less than 5% of the total transformation volume. In conjunction, we recommend using less than 10 ng of DNA. Using a series of DNA amounts is suggested to optimize the protocol and determine which amounts work best for your plasmid before a large amount of transformations are performed.\n\nEnsure you are aware of the dead volume in the containers you are using for the DNA to transform. Dead volumes can be found in the containers section [here](https://developers.transcriptic.com/docs/containers#compatible-container-types). Failure to take into account dead volumes can result in insufficient volume being pipetted.\n[block:api-header]\n{\n  \"type\": \"basic\",\n  \"title\": \"Further reading\"\n}\n[/block]\n[Zymo Research DH5a Cells](https://www.zymoresearch.com/e-coli/chemically-competent-cells/strain-zymo5alpha)\n[Zymo Research 10B Cells](https://www.zymoresearch.com/e-coli/chemically-competent-cells/strain-zymo-10b)\n[Zymo Research JM109 Cells](https://www.zymoresearch.com/e-coli/chemically-competent-cells/strain-jm109)\n[block:callout]\n{\n  \"type\": \"success\",\n  \"body\": \"For further advice on your protocol please ask questions to the community at https://forum.transcriptic.com\",\n  \"title\": \"Check out the forum if you have more questions.\"\n}\n[/block]","excerpt":"","slug":"transform-spread-pick","type":"basic","title":"Transform, Spread, Pick"}

Transform, Spread, Pick


[block:callout] { "type": "warning", "title": "Bacterial Aliquot Expiration", "body": "Please note that bacterial aliquots produced using this protocol will expire and be discarded 3 weeks after their creation. Ordering minipreps, maxipreps, or creating glycerol stocks is recommended within this time frame to preserve successfully transformed bacteria." } [/block] [block:api-header] { "type": "basic", "title": "Description" } [/block] The Transform, Spread, Pick protocol is used for transformation of plasmid DNA into Zymo Mix & Go competent cells followed by colony growth, picking, and OD600 validation. It is often followed by the [Sequencing](doc:sequencing) protocol for sequence verification, and [Plasmid Mini Prep](doc:plasmid-mini-prep) ] or [Plasmid Maxi Prep](doc:plasmid-maxi-prep) protocols to produce aliquots of purified plasmid. [block:api-header] { "type": "basic", "title": "Sample requirements" } [/block] [block:parameters] { "data": { "h-0": "Sample", "h-1": "Explanation", "0-0": "DNA to Transform", "0-1": "An aliquot of plasmid DNA that you wish to transform into chemically competent *E. coli* cells (see below for cell choices)." }, "cols": 2, "rows": 1 } [/block] [block:api-header] { "type": "basic", "title": "Required parameters" } [/block] [block:parameters] { "data": { "h-0": "Parameter", "h-1": "Explanation", "0-0": "Bacteria for Transformation", "0-1": "The bacteria into which you would like to insert your plasmid DNA. All strains are Zymo Mix & Go bacteria designed for quick transformation without the heat shock step. Available strains are Zymo DH5a, Zymo 10B, and Zymo JM109.", "1-0": "Volume to Transform", "2-0": "Colonies", "1-1": "Volume of DNA to transform into competent cells. Must be between 2-5 uL.", "2-1": "Number of colonies to pick per transformation. Must be 1-10.", "3-0": "Recover with SOC Medium at 37C", "3-1": "Adds 400 uL SOC medium to bacteria for a post-transformation recovery at 37C for 50 minutes when selected.", "4-0": "Group Label", "4-1": "The label for one transformation using the specified DNA to Transform. Each group will be transformed using the same specified DNA aliquot(s) and use the same antibiotic for agar plate and growth media.", "5-0": "Antibiotic for Growth Media and Agar Plate", "5-1": "The antibiotic used on the agar plates for spreading the bacteria after transformation, and in the growth media after colony picking. Choices include 100 ug/mL Ampicilin, 50 ug/mL Kanamycin, 100 ug/mL Spectinomycin, 25 ug/mL Chloramphenicol, or No Antibiotic." }, "cols": 2, "rows": 6 } [/block] [block:api-header] { "type": "basic", "title": "Protocol Outputs" } [/block] This protocol outputs new aliquots of bacterial colonies picked into growth media which are stored in your inventory at 4°C. Please note that these bacterial aliquots expire and will be discarded 3 weeks after their creation. Any aliquots of DNA used in the making of these colonies will be returned to the storage condition from which they came. An annotated autopick image is returned showing where on the agar plate colonies were picked. OD600 data is returned for the growth plate to verify colony growth after picking. [block:api-header] { "type": "basic", "title": "Recommended guidelines" } [/block] Zymo Research recommends keeping the added volume of DNA less than 5% of the total transformation volume. In conjunction, we recommend using less than 10 ng of DNA. Using a series of DNA amounts is suggested to optimize the protocol and determine which amounts work best for your plasmid before a large amount of transformations are performed. Ensure you are aware of the dead volume in the containers you are using for the DNA to transform. Dead volumes can be found in the containers section [here](https://developers.transcriptic.com/docs/containers#compatible-container-types). Failure to take into account dead volumes can result in insufficient volume being pipetted. [block:api-header] { "type": "basic", "title": "Further reading" } [/block] [Zymo Research DH5a Cells](https://www.zymoresearch.com/e-coli/chemically-competent-cells/strain-zymo5alpha) [Zymo Research 10B Cells](https://www.zymoresearch.com/e-coli/chemically-competent-cells/strain-zymo-10b) [Zymo Research JM109 Cells](https://www.zymoresearch.com/e-coli/chemically-competent-cells/strain-jm109) [block:callout] { "type": "success", "body": "For further advice on your protocol please ask questions to the community at https://forum.transcriptic.com", "title": "Check out the forum if you have more questions." } [/block]