Alice:LabNotes/2008-7-30: Difference between revisions
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==Experiment: CES22K probe preparation using Jumpstart Taq== | ==Experiment: CES22K probe preparation using Jumpstart Taq== | ||
reaction system 1: x48 | |||
H2O 80.5ul 3864ul | H2O 80.5ul 3864ul | ||
10x Stoffel buffer 10ul 480ul | 10x Stoffel buffer 10ul 480ul | ||
Line 10: | Line 10: | ||
template(CES22K #4) 0.1ul 4.8ul | template(CES22K #4) 0.1ul 4.8ul | ||
Jumpstart Taq 0.6ul 28.8ul | Jumpstart Taq 0.6ul 28.8ul | ||
Reaction system 2: | |||
H2O 80.7ul 3873.6ul | |||
10x Stoffel buffer 10ul 480ul | |||
MgCl2(25mM) 6ul 288ul | |||
dNTP(10mM each, NEB) 2ul 96ul | |||
primer mix (Ribo V6 100uM) 0.2ul 9.6ul | |||
50x SYBG I 0.4ul 19.2ul | |||
template(CES22K #4) 0.1ul 4.8ul | |||
Jumpstart Taq 0.6ul 28.8ul | |||
94c 2min -> 94c 30sec -> 60C 2min -> 72C 1min-> Plate read -> (94c 30sec -> 60C 2min -> 72C 1min)x 20 cycles -> 72C 5min -> 15C hold | 94c 2min -> 94c 30sec -> 60C 2min -> 72C 1min-> Plate read -> (94c 30sec -> 60C 2min -> 72C 1min)x 20 cycles -> 72C 5min -> 15C hold | ||
PCR curve: | |||
[[Image:7-31-08.jpg]] | |||
The reaction system 2 is unsuccessful because the forward primer was not added. | |||
Latest revision as of 21:27, 23 August 2008
Experiment: CES22K probe preparation using Jumpstart Taq[edit]
reaction system 1: x48 H2O 80.5ul 3864ul 10x Stoffel buffer 10ul 480ul MgCl2(25mM) 6ul 288ul dNTP(10mM each, NEB) 2ul 96ul primer mix (Ap6) 0.4ul 19.2ul 50x SYBG I 0.4ul 19.2ul template(CES22K #4) 0.1ul 4.8ul Jumpstart Taq 0.6ul 28.8ul Reaction system 2: H2O 80.7ul 3873.6ul 10x Stoffel buffer 10ul 480ul MgCl2(25mM) 6ul 288ul dNTP(10mM each, NEB) 2ul 96ul primer mix (Ribo V6 100uM) 0.2ul 9.6ul 50x SYBG I 0.4ul 19.2ul template(CES22K #4) 0.1ul 4.8ul Jumpstart Taq 0.6ul 28.8ul 94c 2min -> 94c 30sec -> 60C 2min -> 72C 1min-> Plate read -> (94c 30sec -> 60C 2min -> 72C 1min)x 20 cycles -> 72C 5min -> 15C hold PCR curve:
The reaction system 2 is unsuccessful because the forward primer was not added.