Team:University of Ottawa/30 July 2008

From 2008.igem.org

(Difference between revisions)
(→Today in the Lab)
 
(3 intermediate revisions not shown)
Line 138: Line 138:
:<li> Used PCR cleanup kit to purify digestion products. Measured absorbance to determine concentration.
:<li> Used PCR cleanup kit to purify digestion products. Measured absorbance to determine concentration.
:<li> Concentrations too low for ligation.
:<li> Concentrations too low for ligation.
 +
 +
=='''Matt'''==
 +
:<li> Measured absorbance of pSSA42 gel extraction products from digestion.
 +
 +
=='''Dan'''==
 +
'''Gel Extraction'''
 +
:<li>T123 amplification product was run on a gel and extracted.
 +
:<li>24 gel extraction were performed in total, care was taken to minimize UV exposure (samples were not exposed for longer than 20s)
 +
'''Gel of extracted PCR amplification product'''
 +
:<li> Expected bands were seen, the T123 is good to go.
 +
'''0A and 0B absorbances'''
 +
:<li> absorbances of 0A and 0B indicated very good concentrations
 +
'''Ligation and digestion'''
 +
:<li> Digestion (with SphI and PstI) of T123 0A and 0B were all performed and left overnight

Latest revision as of 14:57, 12 August 2008

Untitled Document

 

 


Contents

Today in the Lab

Tammy

SampleConcentration (ng/μL)V (μL)Total DNA (ng)
pSSA141297903
OB17061020


  • Digestion Step 1 of pSSA14 with PstI
    • Digestion Reaction componentsVol (μl)
    H2O4
    Buffer 31.5
    BSA1.5
    PstI1
    DNA template7
    Total15


  • Digestion Step 1 of OB with PstI
    • Digestion Reaction componentsVol (μl)
    H2O5
    Buffer 31.5
    BSA1.5
    PstI1
    DNA template6
    Total15


  • Digestion Step 2 of pSSA14 with ClaI
    • Digestion Reaction componentsVol (μl)
    H2O9.5
    Buffer 43
    BSA1.5
    ClaI1
    DNA template15
    Total30


  • Digestion Step 2 of OB with ClaI
    • Digestion Reaction componentsVol (μl)
    H2O9.5
    Buffer 43
    BSA1.5
    ClaI1
    DNA template15
    Total30

    Chris

    PCR Confirmation

  • Ran all 5 PCR product samples on a 1% gel for 40 minutes at 80 V
  • Desired band appeared on gel; PCR amplification was successful.
  • Used PCR cleanup kit to purify samples
  • Measured absorbance of PTP2 samples

    • Digestion of PTP2

  • Digested PTP2 with BamHI and XhoI; ran 5 tubes in parallel. Incubated at 37 C for on hour. Ran one water control.
  • Used PCR cleanup kit to purify digestion products. Measured absorbance to determine concentration.
  • Concentrations too low for ligation.

    • Matt

  • Measured absorbance of pSSA42 gel extraction products from digestion.

    • Dan

    Gel Extraction

  • T123 amplification product was run on a gel and extracted.
  • 24 gel extraction were performed in total, care was taken to minimize UV exposure (samples were not exposed for longer than 20s)

    • Gel of extracted PCR amplification product

  • Expected bands were seen, the T123 is good to go.

    • 0A and 0B absorbances

  • absorbances of 0A and 0B indicated very good concentrations

    • Ligation and digestion

  • Digestion (with SphI and PstI) of T123 0A and 0B were all performed and left overnight
    • Retrieved from "http://2008.igem.org/Team:University_of_Ottawa/30_July_2008"