Rui:LabNotes/SingleCell/2011-1-28: Difference between revisions
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>Zsakura2 |
>Zsakura2 |
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===Thoughts=== | ===Thoughts=== | ||
* ExoI is the key point for the unspecific amplification | * ExoI is the key point for the unspecific amplification | ||
* No amplification for cDNA positive control could due to the temperature changes during the procedure by which cDNA was degraded by ExoI. | |||
==Clone screen results (continued)== | ==Clone screen results (continued)== |
Revision as of 20:14, 24 February 2011
Titration Exonuclease I (continued)
Continued with 1.27.11 lab note
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
Thoughts
- ExoI is the key point for the unspecific amplification
- No amplification for cDNA positive control could due to the temperature changes during the procedure by which cDNA was degraded by ExoI.
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???