Carrico:Standard Protocols: Difference between revisions

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QIAquick Gel Extraction Kit Protocol (using a microcentrifuge) - [[:Media:QIAquick Gel Extraction Kit Protocol.pdf|pdf file]]
QIAquick Gel Extraction Kit Protocol (using a microcentrifuge) - [[:Media:QIAquick Gel Extraction Kit Protocol.pdf|pdf file]]
===How to determine DNA template concentration (and amount)===
#Find absorbance (OD) by:
##In a micro cell, pipette 50uL water
##BLANK at 260nm (remember to turn both vis & UV lamps on and wait 5 minutes)
##Pipette out 5uL water
##Pipette in 5uL DNA template
##Place micro cell in spectrophotometer & READ (remember light pathway for placement)
#Clean micro cell by washing with 2mL water followed by 2mL methanol/ethanol from wash bottles
#Go to [http://www.pubquizhelp.34sp.com/other/dnacalculator.html/ DNA Concentration Calculator]


=="How to" for Vector NTI==
=="How to" for Vector NTI==

Revision as of 14:21, 22 September 2008


Carrico Lab at Stony Brook


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QIAquick Gel Extraction Kit Protocol (using a microcentrifuge) - pdf file

How to determine DNA template concentration (and amount)

  1. Find absorbance (OD) by:
    1. In a micro cell, pipette 50uL water
    2. BLANK at 260nm (remember to turn both vis & UV lamps on and wait 5 minutes)
    3. Pipette out 5uL water
    4. Pipette in 5uL DNA template
    5. Place micro cell in spectrophotometer & READ (remember light pathway for placement)
  2. Clean micro cell by washing with 2mL water followed by 2mL methanol/ethanol from wash bottles
  3. Go to DNA Concentration Calculator

"How to" for Vector NTI

How to create a new plasmid

  1. Open the Vector NTI Explorer program
  2. Click on the "New" icon
  3. Fill-in "Name"
  4. Under the "Sequence" tab, click on "Edit Sequence"
  5. Paste-in the sequence
  6. Select "OK"
  7. Double-click on newly created plasmid

How to create a gene in a plasmid

  1. Highlight the plasmid you would like edited
  2. Select "Edit" --> "New" --> "Add feature"
  3. Select the name of the plasmid
  4. Select "Save as"

How to change nucleotides

  1. Highlight bases to be changed
  2. Select "Edit" --> "New" --> "Replace"
  3. "Save As" a different name