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Originally posted by zhangzhuang at 2010-06-28 15:47:42:
´ó¼ÒË­×ö¹ýÓ«¹â¶¨Á¿PCRѽ£¿
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¹þ¹þ£¬ÒÔǰ£¬»¹ÕæÊÇΪÁ˸øÀÏÍâ½²¿Î£¬×÷ÁËÒ»¸öStepone plus ²Ù×÷²½Öè ppt£¬²»¹ýÏÖÔÚ²»ÖªµÀÈ¥ÄÇÀïÁË£¬ÄãÓÐʲôÎÊÌ⣬¾ÍÖ±½Ó˵¾ÍÊÇÁË£¬
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1.  Open the ABI StepOnePlus software on the desktop of the computer that is connected to
the ABI StepOnePlus system.

2.  Select Advanced Setup  

3.  Define: Experiment Properties

a.  Label Your Experiment
i.  Type In Experiment Name
ii.  Type In Barcode, User Name, Comments (Optional)
b.  Select Instrument
i.  StepOnePlus Instrument (96 wells)
c.  Select Experiment Type
i.  Quantitation-Standard Curve
d.  Select Reagents
i.  SYBR Green
e.  Select Ramp Speed
i.  Standard (~2 hours to complete)

4.  Click Plate Setup (on left)
a.  Click Assign Targets and Samples Tab
i.  Highlight the entire plate
ii.  Check the box next to Target 1 under Assign targets to the selected wells
iii.  Verify all wells in Plate Layout view have the ¡°U¡± symbol (¡°U¡± =
unknown)

5.  Click Run Method (This setting should default to run protocol with melting curve.)
a.  Verify Data Capture icon is present at:
i.  Cycling Stage: 60¡ãC (1 minute step)
ii.  Melting Curve Stage: During ramp from 60¡ãC to 95¡ãC
b.  Set Reaction Volume to 25 uL
c.  Verify Number of Cycles is set to 40

6.  Click Start Run

[ Last edited by reasonspare on 2010-6-29 at 04:52 ]
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