DNase Protocol

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Contents

Overview

Protocol for the Ambion DNA-free DNase kit. This uses a DNase inactivating reagent and thus avoids the DNase heat inactivation step that makes me nervous.

Materials

List reagents, supplies and equipment necessary to perform the protocol here. For those materials which have their own OWW pages, link to that page. Alternatively, links to the suppliers' page on that material are also appropriate.

  • supply 1 (i.e. tubes of a certain size? spreaders?)
  • reagent 1
  • X μL reagent 2
    • component A (reagent 2 is made up of multiple components)
    • component B
  • equipment 1
  • equipment 2

Procedure

1) In order to DNase 20ug of RNA in a 100ul reaction, mixup the following:

    • 10X DNase buffer:10ul
    • dH20:78ul
    • DNase:2ul
    • RNA(1ug/ul):10ul
  1. Step 2
    • Step 2 has some additional information that goes with it. i.e. Keep at 4°C.
  2. Step 3
    1. Step 3 has multiple sub-steps within it.
    2. Enumerate each of those.

Notes

  1. List troubleshooting tips here.
  2. You can also link to FAQs/tips provided by other sources such as the manufacturer or other websites.
  3. Anecdotal observations that might be of use to others can also be posted here.

Please sign your name to your note by adding '''*~~~~''': to the beginning of your tip.

References

Relevant papers and books

  1. Goldbeter A and Koshland DE Jr. . pmid:6947258. PubMed HubMed [Goldbeter-PNAS-1981]
  2. JACOB F and MONOD J. . pmid:13718526. PubMed HubMed [Jacob-JMB-1961]
  3. Mark Ptashne. A genetic switch. Cold Spring Harbor, N.Y.: Cold Spring Harbor Laboratory Press, 2004. isbn:0879697164. [Ptashne-Genetic-Switch]
All Medline abstracts: PubMed HubMed

Contact

  • Who has experience with this protocol?

or instead, discuss this protocol.


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