Rui:LabNotes/SingleCell/2011-1-28

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Revision as of 20:13, 24 February 2011 by >Zsakura2 (→‎Clone screen results (continued))
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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
  • Thoughts???