Matt:LabNotes/Qubit Protocol: Difference between revisions

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(Created page with "==Qubit Protocol== *Taken from Brandon's protocols Qubit can be used for dsDNA, ssDNA, dsDNA BR and RNA quantification. 1. make Qubit mastermix (dye and buffer) per sample...")
 
>Mzcai
 
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==Qubit Protocol==
==Qubit Protocol==
*Taken from Brandon's protocols


Qubit can be used for dsDNA, ssDNA, dsDNA BR and RNA quantification.
Qubit can be used for dsDNA, ssDNA, dsDNA BR and RNA quantification.
(high sensitivity for 2ng/ul - 100ng/ul)




1. make Qubit mastermix (dye and buffer)
1. make Qubit mastermix (dye and buffer)
  per sample:
  per sample:
  1 uL 200X dye (Component A, Quant-IT RNA reagent (200X))
  1 uL 200X dye (Component A, Quant-IT reagent (200X))
  199 uL Buffer (Component B, Quant-IT RNA buffer)
  199 uL Buffer (Component B, Quant-IT buffer)





Latest revision as of 23:21, 30 April 2013

Qubit Protocol[edit]

Qubit can be used for dsDNA, ssDNA, dsDNA BR and RNA quantification. (high sensitivity for 2ng/ul - 100ng/ul)


1. make Qubit mastermix (dye and buffer)

per sample:
1 uL 200X dye (Component A, Quant-IT reagent (200X))
199 uL Buffer (Component B, Quant-IT buffer)


2. Prepare samples and controls in qubit assay tubes (invitrogen)

a. add 10 uL RNA Standard #1, 0 ng (component C) to 190 uL mastermix

b. add 10 uL RNA Standard #2, 10 ng (component D) to 190 uL mastermix

c. add 1-10 uL RNA to 190-199 uL mastermix for each sample


3. If needed, let sit in dark for at least 2 minutes after each sample is added


4. Start "Qubit data logger" on the computer. Push start, button will turn green with "port open"


5. On Qubit device, push "Home" to start qubit, then push "Go" and select the assay. Then do calibration with the standards by putting the qubit tube with a standard into the qubit device, be sure to mix before hand!

Standard 1 = 0  ng (component C)
Standard 2 = 10 ng (component D)


6. After calibration with standards, samples can now be run. Be sure to mix before hand! Select "calculate concentration" after each sample and enter the input sample volume (1-10 uL) to get concentration in ug/mL.


7. Can export data and save.