Protocols/Consensus template

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==Procedure==
==Procedure==
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A step by step guide to the experimental procedure.
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1. Place Glass plates in casting apparatus
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2. Add together the following to make 5ml of gel (0.75mm spacers)
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If you find it helpfull you could use the following icons to highlight important parts:
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3ml water
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1.5ml 29:1 acrylamide solution (40% w/v)
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[[Image:Difficult step.png]] icon to highlight difficult steps
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500µl10X TBE solution
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35µl Ammonium Persulfate (10%w/v)
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[[Image:Critical step.png]] warn about critical steps
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2µl TEMED
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3. Pipet mix between casting plates using 5ml pipetor
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[[Image:Time required.png]] help people plan how long steps will take
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4. Insert comb and allow to cure for 30 minutes
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5. Mix the following for all DNA samples including the ladder
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[[Image:Pause point.png]] where protocol can be interrupted
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10 DNA
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1ul SYBR green  (1:100 Dilution in DMSO)
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[[Image:Optional step.png]] steps that can be omitted or included
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2ul Bromophenol Blue
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6. Remove comb, insert gel into electrophoresis chamber and add the necessary amounts of 1X TBE
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7. Add DNA mix to wells. 
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8. Apply 80 volts and run for approximately 60 minutes
==Critical steps==
==Critical steps==

Revision as of 14:36, 17 June 2009

This page is a template and should not be edited.
Click here, copy the source, and paste it into your page.



Interested in posting a new template for a consensus protocol on OpenWetWare? Here is a template to help you do so.

Click the Image:EditButton.jpg tab at the top and copy everything below this line. Paste it into your new protocol page. Then replace the text in this page with your own protocol. Feel free to add or delete sections as appropriate.

Contents

Curators

Add the primary curators here (including their contact info). For instance,

James Hadfield, CRUK Cambridge Research Institute, Robinson Way, Cambridge CB2 0RE. Tel: +44 (0)1223 404250; Fax: +44 (0)1223 404208; email: jamesdothadfieldatcancerdoyorgdotuk.

Anyone should feel free to add themselves as a curator for this consensus protocol. You do not need to be a curator in order to contribute. The OpenWetWare community is currently discussing the idea of protocol curators. Please contribute.

Abstract

Replace this sentence with a brief description of the protocol and its goal.

Materials

List everything necessary to perform the protocol here. Include all information about suppliers, ordering details, etc. Links to the suppliers' page on that material are also appropriate and encouraged. Please be aware that users of this protocol may not be working in the same country as you.

Reagents

Biological resources e.g. cell lines, buffers (link to a method for making them), enzymes, chemicals, kits, etc.

Equipment

Any equipment used to perform the protocol (link to a method for using them).

Procedure

1. Place Glass plates in casting apparatus 2. Add together the following to make 5ml of gel (0.75mm spacers) 3ml water 1.5ml 29:1 acrylamide solution (40% w/v) 500µl10X TBE solution 35µl Ammonium Persulfate (10%w/v) 2µl TEMED 3. Pipet mix between casting plates using 5ml pipetor 4. Insert comb and allow to cure for 30 minutes 5. Mix the following for all DNA samples including the ladder 10 DNA 1ul SYBR green (1:100 Dilution in DMSO) 2ul Bromophenol Blue

6. Remove comb, insert gel into electrophoresis chamber and add the necessary amounts of 1X TBE 7. Add DNA mix to wells. 8. Apply 80 volts and run for approximately 60 minutes

Critical steps

Referenced from the main protocol, a more thorough explanation of particularly important steps in the protocol.

Troubleshooting

Referenced from the main protocol, an explanation of what can cause things to go wrong with the protocol.

Notes

Referenced from the main protocol, any comments about the protocol should be made here; i.e. how it was developed. Any comments added should be signed (by adding *'''~~~~''': in front) and explained. Links to FAQs/tips provided by other sources such as the manufacturer or other websites would be best made here.
Anecdotal observations that might be of use to others can also be posted here; e.g. 'my cells were still floating'.

It might also be good to add an image to show the workflow and timescales for experiment planning.

Acknowledgments

Acnkowledge any help you had in development, testing, writing this protocol.

References

See OpenWetWare:Biblio for information on how to reference within a wiki.

Specific Protocols

Add links to all the OWW protocols that have been used in making the consensus.

Discussion

You can discuss this protocol.

Tag this page with categories to allow easier indexing and searching. See Categories for information on existing categories.

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