Protocols/Template: Difference between revisions

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{|style="width: 48em; background: #FF6677;"
| align="center"|'''This page is a template and should not be edited.'''<br><span style="font-size:90%">Click [http://openwetware.org/index.php?title={{PAGENAME}}&action=edit here], copy the source, and paste it into your page.</span>
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'''Interested in posting a protocol on OpenWetWare?  Here is a template to help you do so.'''   
'''Interested in posting a protocol on OpenWetWare?  Here is a template to help you do so.'''   


'''Click the edit tab and copy everything below this line.  Paste it into your new protocol page.'''
'''Click the view source tab 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.'''
[[Category:Template]]
<!-- COPY EVERYHING BELOW HERE TO START YOUR OWN PROTOCOL!  -->


==Overview==
==Overview==
Line 8: Line 20:


==Materials==
==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?)
For a 50 μL PCR reaction:
*reagent 1
 
*X &mu;L reagent 2
* 35 μL H<sub>2</sub>O
**component A (reagent 2 is made up of multiple components)
* 5 μL 10X PCR buffer
**component B
* 5 μL 2mM dNTPs (each)
*equipment 1
* 1.5 μL 50mM MgCl<sub>2</sub>
*equipment 2
* 1 μL 50μM sense primer
* 1 μL 50μM antisense primer
* 1 μL 5nM DNA template
* 0.5 μL TAQ DNA polyermerase


==Procedure==
==Procedure==
#Step 1
# In a PCR tube, mix the components on ice in the order they are listed above.
#Step 2
# Perform thermocycling program
#*Step 2 has some additional information that goes with it.  i.e. Keep at 4&deg;C.
## 95 °C 5 min
#Step 3
## 95 °C 30 s
##Step 3 has multiple sub-steps within it.
## T<sub>H</sub> 30 s
##Enumerate each of those.
## 72 °C 1 min for each 1 kb PCR product
## Repeat steps 2-4 a total of 12-36 times (24 is standard).
## 72 °C 5 min
## 12 °C hold


==Notes==
==Notes==
#List troubleshooting tips here.
Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!
#List troubleshooting tips here.
#You can also link to FAQs/tips provided by other sources such as the manufacturer or other websites.
#You can also link to FAQs/tips provided by other sources such as the manufacturer or other websites.
#Anecdotal observations that might be of use to others can also be posted here.
#Anecdotal observations that might be of use to others can also be posted here. 
 
Please sign your name to your note by adding <font face="courier"><nowiki>'''*~~~~''':</nowiki></font> to the beginning of your tip.


==References==
==References==
If this protocol has papers or books associated with it, list those references here. Use the [[OpenWetWare:Biblio]] extension to standardize formatting and provide relevant PubMed links.
'''Relevant papers and books'''
<!-- If this protocol has papers or books associated with it, list those references here.-->
<!-- See the [[OpenWetWare:Biblio]] page for more information, but this format doesn't work currently.
<biblio>
#Goldbeter-PNAS-1981 pmid=6947258
#Jacob-JMB-1961 pmid=13718526
#Ptashne-Genetic-Switch isbn=0879697164
</biblio>-->
<!-- Try the [[Template:FormatRef|FormatRef template]]-->
#{{FormatRef|Goldbeter, A and Koshland, DE|1981| |Proc Natl Acad Sci U S A 78(11)|6840-4| }} PMID 6947258
#{{FormatRef|Jacob, F and Monod, J J|1961| |Mol Biol 3(3)|318-56| }} PMID 13718526
#{{FormatRef|Ptashne, M|2004|Genetic Switch: Phage Lambda Revisited| | |Cold Spring Harbor Laboratory Press}} ISBN 0879697164
 
==Contact==
*Who has experience with this protocol?
 
or instead, [[Talk:{{PAGENAME}}|discuss this protocol]].
 
<!-- You can tag this protocol with various categories.  See the [[Categories]] page for more information. -->
 
<!-- Move the relevant categories above this line to tag your protocol with the label
[[Category:Protocol]]
 
[[Category:Needs attention]]
[[Category:In vitro]]
 
[[Category:In vivo]]
 
[[Category:In silico]]
 
[[Category:DNA]]
 
[[Category:RNA]]
 
[[Category:Protein]]
 
[[Category:Chemical]]
 
[[Category:Escherichia coli]]
 
[[Category:Yeast]]
-->

Latest revision as of 14:27, 18 June 2012

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 protocol on OpenWetWare? Here is a template to help you do so.

Click the view source tab 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.

Overview

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

Materials

For a 50 μL PCR reaction:

  • 35 μL H2O
  • 5 μL 10X PCR buffer
  • 5 μL 2mM dNTPs (each)
  • 1.5 μL 50mM MgCl2
  • 1 μL 50μM sense primer
  • 1 μL 50μM antisense primer
  • 1 μL 5nM DNA template
  • 0.5 μL TAQ DNA polyermerase

Procedure

  1. In a PCR tube, mix the components on ice in the order they are listed above.
  2. Perform thermocycling program
    1. 95 °C 5 min
    2. 95 °C 30 s
    3. TH 30 s
    4. 72 °C 1 min for each 1 kb PCR product
    5. Repeat steps 2-4 a total of 12-36 times (24 is standard).
    6. 72 °C 5 min
    7. 12 °C hold

Notes

Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!

  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 (1981) - Proc Natl Acad Sci U S A 78(11) 6840-4 PMID 6947258
  2. Jacob, F and Monod, J J (1961) - Mol Biol 3(3) 318-56 PMID 13718526
  3. Ptashne, M (2004) Genetic Switch: Phage Lambda Revisited - Cold Spring Harbor Laboratory Press ISBN 0879697164

Contact

  • Who has experience with this protocol?

or instead, discuss this protocol.


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