Author checklist

  1. Please provide a list of keywords that can be used as search terms.
  2. Please indicate the corresponding author and provide email address.

Abstract

  1. Please include a sentence to explain how long the protocol takes to complete.
  2. Please briefly explain how the protocol compares to alternative methods.

Introduction

  1. Please start by introducing the methods used in the protocol, including references to papers that catalogue their development and previous use. Please compare your approach to alternative methods and fully describe the potential applications and limitations of the method.
  2. In the ‘Experimental Design’ section, please provide all information about the design of experiments that readers would need to adapt the protocol to apply to their own experimental situation. You should also discuss the controls necessary for the protocol.
  3. We encourage readers to adhere to the ARRIVE guidelines for the reporting of animal experiments(PLoS Bio 8(6), e1000412,2010). Please include a CAUTION note in the Materials and Procedure sections (and relevant figure legends), outlining that appropriate institutional regulatory board guidelines must be followed and explain the permission that was obtained.
  4. The description of microarray experiments should adhere to the standards outlined in the MIAME guidelines (MGED web site).Please ensure relevant aspects are described in the Experimental Design and Procedure sections.

Materials

  1. Please list every reagent and piece of equipment individually (except for basic lab equipment), with supplier details and catalogue numbers.
  2. Please add a CAUTIONnote to any potentially harmful items and explain the danger plus precautions that should be taken when handling them.
  3. Please add a CRITICAL note if it is crucial that a particular reagent/supplier is used and explain why.
  4. For every item, please indicate whether it should be made up fresh or can be stored and, if so, under which conditions and for how long.
  5. For every %, please state whether this iswt/vol or vol/vol.

Procedure

  1. The Procedure must take the form of a continuously numbered set of steps (i.e. use 1, 2, 3, 4; not 1.1, 1.2 etc.), with numbering continuing from subsection to subsection. There can only be one level of subheading.
  2. If there are options of how to proceed at a particular step, options A, B, C etc. can be used within that step, and within these options, different steps should be numbered using roman numerals. Please indicate if any steps are optional.
  3. If there are any additional procedures that do not fit into the continuous flow of numbered steps, these can be put into boxes and referred to from the Procedure.
  4. Our format does not allow for any additional information in the Procedure that is not within a numbered step. Please place all additional information in the Introduction or Anticipated Results.
  5. Please write the Procedure steps in the active tense. For example, “Add 50ml of solution X.”
  6. If possible, please add timing information to all Procedure subheadings.
  7. All Troubleshooting information should be removed from the Procedure and combined to form an independent Troubleshooting section at the end of the Procedure.
  8. Please give all centrifugation speeds in xg.
  9. Please state what room temperature is in your laboratory the first time you use the term.

Timing

  1. Please include a Timing section.
  2. Where possible, please add corresponding Procedure step numbers to the entries in this section.

Troubleshooting

  1. Please include a Troubleshooting section. This should preferably take the form of a table with four columns, entitled ‘Step’, ‘Problem’, ‘Possible reason’ and ‘Possible solution’.
  2. Please add ?Troubleshootingafter each step in the Procedure for which you have given troubleshooting guidance in the Troubleshooting Table.

Anticipated results

  1. Please include an Anticipated Results section to demonstrate the types of results that can be expected from the protocol and explain how they can be interpreted. Figures showing examples of good or poor quality results can also be referred to from this section.
  2. Ensure data is fully explained (n values given, error bars defined, etc.). Graphs showing individual data points are often preferable to those showing means and error bars. If there are fewer than 6 data points, individual data points must be provided. Please ensure any statistics are applied appropriately (

Figures/Tables/Boxes

  1. All Figures/Tables/Boxes must be cited in the main text and should be numbered in the order in which the citations appear in the text.
  2. Please submit figures individually in one of our accepted file formats (preferably eps or tif).
  3. Please provide a title and figure legend for each figure. Individual panels cannot be cited from the title. Figures legends should fully explain the corresponding figure. Please include legends in the main manuscript.
  4. If Figures have been published previously, please refer to the original source in the legend.
  5. Please include a scale bar if appropriate and indicate its length in the corresponding figure legend.
  6. The colour yellow can be difficult to distinguish; please consider replacing it with another color.
  7. Please avoid the use of red and green to circumvent issues with color blindness.
  8. Please avoid the use of colored backgrounds in figures/flowcharts.
  9. Please remove any boxes around your graphs/histograms/plots, unless x, y and z axes are used.
  10. Please remove boxes around figures/figure panels unless there is a specific reason for the box.
  11. Please check the size (total dimensions and number of panels) of each figurefits on a single page of PDF (including legend). For large figures, consider splitting into smaller separate figures.
  12. Please consider the relative importance of panels. If an image or panel within a figure is particularly important, please let us know (otherwise, panels will be about the same size by default).
  13. Please do not use patterned fills in bar or pie charts. Fills must be solid colors only.
  14. Please do not put tables inside figures –data should be presented in a separate, numbered table.
  15. Axis labels should be clear, accurate, and informative (including relevant units)
  16. Please include origins and intervals on axes – most graphs will have zero origins and regular intervals
  17. Avoid subpanels unless absolutely crucial
  18. All fonts should be sans serif, and bold/italic text should be avoided except where scientifically necessary (e.g., italics for variables, bold for compound numbers)

CFI

  1. Please include the appropriate competing financial interest statement: either “The authors declare that they have no competing financial interests.”, or “The authors declare competing financial interests (see the HTML version of this article for details).”
  2. If relevant, please then provide an explanation of what these potential competing interests consist of.

Acknowledgements

  1. Grants/awards should be mentioned here.

Author contributions statement

  1. Please include a statement that specifies the contribution of every author (see in your preferred wording.

Supplementary information

  1. Each piece of Supplementary Information (SI) should have a title and be cited in order in the text. Where appropriate, legends should be included that refer to individual panels and/or symbols.
  2. Please list all SI, including the types, titles and legends, at the end of your Manuscript File.
  3. Please place any supplementary figures and their associated titles and legends in our Integrated Supplementary Figures template ( For any PDF-able SI that are not integrated figures (e.g. tables, notes, manual, methods, etc), please create a single, combined PDF.
  4. Please submit any SI that are not PDF-able (e.g. videos, podcasts, Excel docs, etc) separately, but remember to include SI titles and legends at the end of the manuscript.