Matt:LabNotes/2014-10-6: Difference between revisions

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>Mzcai
(Created page with "==Testing Agi26k0gap + suppressor Oligos & Agi26k20gap== ===Sample=== *Agi26k0gap Control: PGP1F Fix 5.19.14 S2 *Agi26k0gap + suppressors: PGP1F Fix 5.19.14 S3 *Agi26k20gap: ...")
 
>Mzcai
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| 10X Ampligase Buffer||10
| 10X Ampligase Buffer||10
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| [[Matt:LabNotes/2013-6-4 | Agi26k20gap]]||77
| [[Matt:LabNotes/2013-6-4 | Agi26k20gap]]||70
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| 25mM dNTP||1
| 25mM dNTP||1
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| 10 U/ul KlenTaq||2
| 10 U/ul KlenTaq||2
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| H2O||0
| H2O||7
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| Total||100
| Total||100

Revision as of 22:00, 6 October 2014

Testing Agi26k0gap + suppressor Oligos & Agi26k20gap

Sample

  • Agi26k0gap Control: PGP1F Fix 5.19.14 S2
  • Agi26k0gap + suppressors: PGP1F Fix 5.19.14 S3
  • Agi26k20gap: Fix 10.1.14

Procedure

  • Agi26k0gap Control
Components Volume
5 U/ul Ampligase 10
10X Ampligase Buffer 10
Agi26k0gap 35
H2O 45
Total 100
  • Agi26k0gap + suppressors
Components Volume
5 U/ul Ampligase 10
10X Ampligase Buffer 10
Agi26k0gap 35
600 nM (100nM each) supp_Mix 5
H2O 40
Total 100
  • Agi26k20gap
Components Volume
5 U/ul Ampligase 10
10X Ampligase Buffer 10
Agi26k20gap 70
25mM dNTP 1
10 U/ul KlenTaq 2
H2O 7
Total 100
  • Incubate at 37C for 30 min
  • Incubate at 60C for 24 hrs