Kun:LabNotes/ASE/2007-10-29: Difference between revisions

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*Starting from ethanol precipitated DNAs made at [[Kun:LabNotes/ASE/2007-10-18 |10/18/2007]].<br>
*Starting from ethanol precipitated DNAs made at [[Kun:LabNotes/ASE/2007-10-18 |10/18/2007]].<br>
*Pool two tubes of the same amplicons into ~100ul in total, add 11ul 10x Lambda Exo buffer, 4ul 5U/ul Lambda exonuclease. Incubate @ 37C for 2 hours, check 2ul of the products in both 8% TBE gels and 6% TBU gels.
*Pool two tubes of the same amplicons into ~100ul in total, add 11ul 10x Lambda Exo buffer, 4ul 5U/ul Lambda exonuclease. Incubate @ 37C for 2 hours, check 2ul of the products in both 8% TBE gels and 6% TBU gels.
 
                      8% TBE gel                            6% TBU gel
         [[image:2007-10-29-PCR-Lambda_exo-TBE.jpg]] [[image:2007-10-29-PCR-Lambda_exo-TBU.jpg]]
         [[image:2007-10-29-PCR-Lambda_exo-TBE.jpg]] [[image:2007-10-29-PCR-Lambda_exo-TBU.jpg]]


*I saw sharp bands in the non-denaturing gel, suggesting there are still a lot of dsDNA. I decided to incubate the reactions for another 4 hours.
*I saw sharp bands in the non-denaturing gel, suggesting there are still a lot of dsDNA. I decided to incubate the reactions for another 4 hours.

Revision as of 01:07, 30 October 2007

Exp1: Probe prep (cont.)

  • Starting from ethanol precipitated DNAs made at 10/18/2007.
  • Pool two tubes of the same amplicons into ~100ul in total, add 11ul 10x Lambda Exo buffer, 4ul 5U/ul Lambda exonuclease. Incubate @ 37C for 2 hours, check 2ul of the products in both 8% TBE gels and 6% TBU gels.
                      8% TBE gel                            6% TBU gel
        File:2007-10-29-PCR-Lambda exo-TBE.jpg File:2007-10-29-PCR-Lambda exo-TBU.jpg
  • I saw sharp bands in the non-denaturing gel, suggesting there are still a lot of dsDNA. I decided to incubate the reactions for another 4 hours.