Alice:LabNotes/2008-7-30

From ZhangLabWiki
Revision as of 21:27, 23 August 2008 by >Zsakura2
(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)
Jump to navigation Jump to search

Experiment: CES22K probe preparation using Jumpstart Taq

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:

File:7-31-08.jpg

The reaction system 2 is unsuccessful because the forward primer was not added.