Zebrafish Care and Experimental Techniques: Real Time PCR

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Materials:
-primers (24 and 25 are for slitrk 3)
-cDNA (40 hour/90 hour)
-10mM dNTP (nucleotides)
-buffer 5x
-water
-enzyme (cooler in freezer)

Combine in a PCR tube (50 microliters of solution):
1. 35.5 uL of water
2. 10uL 5x buffer
2. 1uL of JR 24
3. 1uL of JR 25
4. 1uL of dNTP
5.1uL of cDNA
6. 0.5 uL of Fusion hot start enzyme

Add water to the PCR tube first. Make sure you use the PCR tubes because they fit in the machine.
After all contents have been added, flick PCR tube and centrifuge for 4 seconds.

To Run the PCR Machine:
The power switch is on the back. The lid will click, and then the wheel should be twisted until you feel resistance.
Program: JRRTPCR
enable heated lid
The reaction takes several hours. The machine with have a block temp of 4 degrees when done.
Remember to turn off the machine when finished using it.

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