Haynes Lab:Notebook/SynBERC Scholars 2013/2014/03/17: Difference between revisions

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Line 26: Line 26:
| Reagent || 1 || 2 || 3 || 4 || 5 || 6
| Reagent || 1 || 2 || 3 || 4 || 5 || 6
|-
|-
| Genomic DNA (500 ng) || 2.0 μL || 2.0 || 2.0 || 1.0 || 1.0 || 1.0
| Genomic DNA (~500 ng) || 2.0 μL || 2.0 || 2.0 || 1.0 || 1.0 || 1.0
|-
|-
| Rest. enzyme || 0 || 2.0 || 2.0 || 0 || 2.0 || 2.0
| Rest. enzyme || 0 || 2.0 || 2.0 || 0 || 2.0 || 2.0
Line 48: Line 48:
{| {{table}}
{| {{table}}
|-
|-
| Reagent || Kozak || Luc14 || 3
| Reagent || Kozak || Luc14
|-
|-
| Genomic DNA (500 ng) || 2.0 μL || 2.0 || 2.0 || 1.0 || 1.0 || 1.0
| DNA || 0.5 μL || 0.5
|-
|-
| Rest. enzyme || 0 || 2.0 || 2.0 || 0 || 2.0 || 2.0
| P0001 || 1.0 || ---
|-
| P0002 || 1.0 || ---
|-
|-
| 10x buffer || 2.0 || 2.0 || 2.0 || 2.0 || 2.0 || 2.0
| mixed primers || --- || 1.0
|-
|-
| dH<sub>2</sub>O || 16.0 || 14.0 || 14.0 || 17.0 || 15.0 || 15.0
| dH<sub>2</sub>O || 16.0 || 14.0 || 14.0 || 17.0 || 15.0 || 15.0

Revision as of 10:43, 21 March 2014

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03/17/14

  • Luc 14 genomic DNA test - restriction digest and gel
  • Luc 14 genomic DNA clean-up & PCR


Luc 14 genomic DNA test - restriction digest and gel

  • Check genomic DNA on a gel, with or without restriction digestion
  1. Luc 14 HEK - No enzyme
  2. Luc 14 HEK - EcoRI (fast digest)
  3. Luc 14 HEK - PstI (fast digest)
  4. U2OS plain - No enzyme
  5. U2OS plain - EcoRI (fast digest)
  6. U2OS plain - PstI (fast digest)
Reagent 1 2 3 4 5 6
Genomic DNA (~500 ng) 2.0 μL 2.0 2.0 1.0 1.0 1.0
Rest. enzyme 0 2.0 2.0 0 2.0 2.0
10x buffer 2.0 2.0 2.0 2.0 2.0 2.0
dH2O 16.0 14.0 14.0 17.0 15.0 15.0
  20.0 20.0 20.0 20.0 20.0 20.0


Luc 14 genomic DNA clean-up & PCR

  • Clean up 1/2 of the Luc genomic DNA (50 μL) on a Zymo clean and concentrator column
    • 50 μL ~ 4340 ng
    • Eluted with 20 μL dH2O ~ 217 ng/μL


Reagent Kozak Luc14
DNA 0.5 μL 0.5
P0001 1.0 ---
P0002 1.0 ---
mixed primers --- 1.0
dH2O 16.0 14.0 14.0 17.0 15.0 15.0
  20.0 20.0 20.0 20.0 20.0 20.0



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