Rui:LabNotes/SingleCell/2011-1-28: Difference between revisions

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==Clone screen results (continued)==
==Clone screen results (continued)==
 
===Continue with 1.27.11 lab note===
[http://genome-tech.ucsd.edu/LabNotes/index.php?title=Rui:LabNotes/SingleCell/2011-1-27]
===Gel check===
  *Upper part: NTC-ES; NTC-EB
  *Upper part: NTC-ES; NTC-EB
  *Lower part: 10pg-EB; 500pg-EB
  *Lower part: 10pg-EB; 500pg-EB

Revision as of 20:09, 24 February 2011

Titration Exonuclease I (continued)

Continued with 1.27.11 lab note

[1]

Results

*Each samples (tube1 to 8) have 4 aliquots for PCR, 40ul/aliquotes
*4ul PCR products from one aliquot per sample was checked by 6% TBE-PAGE
*30min running time 
Sample arrangement 
        Template         P1 primer   SSIII    Exo buffer    ExoI       Final
Tube1   water            0.125ul     0.33ul   1* in 1ul     0.5U       0.16* in 6ul
Tube2   water            0.125ul     0.33ul   1* in 1ul     2U         0.16* in 6ul
Tube3   water            0.125ul     0.33ul   3* in 1ul     4U         0.5* in 6ul
Tube4   water            0.125ul     0.33ul   6* in 1ul     8U         1* in 6ul
Tube5   0.1pg 6-EB cDNA                       1* in 1ul     0.5U       0.16* in 6ul       
Tube6   0.1pg 6-EB cDNA                       1* in 1ul     2U         0.16* in 6ul
Tube7   0.1pg 6-EB cDNA                       3* in 1ul     4U         0.5* in 6ul
Tube8   0.1pg 6-EB cDNA                       6* in 1ul     8U         1* in 6ul

File:Exo I.jpg

Thoughts

  • ExoI is the key point for the unspecific amplification


Clone screen results (continued)

Continue with 1.27.11 lab note

[2]

Gel check

*Upper part: NTC-ES; NTC-EB
*Lower part: 10pg-EB; 500pg-EB
*5ul out of 50ul PCR products
File:Clone screen.jpg