Sam:LabNotes/Microbiome/2009-3-11: Difference between revisions

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>Sam Chiang
(New page: ='''PCR confirmation for human MDA amplicons & Q-solution test on Qiagen MPCR kit'''= ==Objectives== *Compare if the Q-solution can improve the results using Qiagen MPCR kit *Pick up mor...)
 
>Sam Chiang
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*Pick up more candidates of amplicons to be tested using the Taq2X MPCR with set-up condition
*Pick up more candidates of amplicons to be tested using the Taq2X MPCR with set-up condition


==Sample & Materials==
 
==Samples & Materials==
 
*Primers:
**Blue primers mix: Y-1(122), 21-1(200), 1-1(270), 5-1(413), 22-1(757)
**Yello primers mix: 11-1(131), 10-1(214), 15-1(297), 17-1(419), 16-1(806)
 
*Templates
**For all Qiagen MPCR (QMPCR) tests: 5 uL of 1 ng/uL human gDNA
**For TaqPol2X MPCR (TMPCR) tests: 1 uL of Amplicon candidates picked up from human MDA (03/10/09): B10, E9, E10, F9, F10, H8, H9, H10
 
 
==Procedures==
 
*QMPCR - no Q-solution
                            1 rxn     
    -----------------------------
    RNAse-free H2O          15.0
    Template (1 ng/uL)        5.0 
    Qiagen kit enzyme (2X)  25.0
    -----------------------------
                              45.0 (uL)
    Add 5 uL primer Bule / Yellow
 
*QMPCR - with Q-solution
                            1 rxn
    -----------------------------
    RNAse-free H2O          10.0
    Q-solution                5.0
    Template (1 ng/uL)        5.0 
    Qiagen kit enzyme (2X)  25.0
    -----------------------------
                              45.0 (uL)
    Add 5 uL primer Bule / Yellow
 
*TMPCR
                            1 rxn     
    -----------------------------
    RNAse-free H2O          19.0
    Template (1 ng/uL)        1.0 
    NEB Taq pol.(2X)        25.0
    -----------------------------
                              45.0 (uL)
    Add 5 uL primer Bule

Revision as of 19:07, 12 March 2009

PCR confirmation for human MDA amplicons & Q-solution test on Qiagen MPCR kit

Objectives

  • Compare if the Q-solution can improve the results using Qiagen MPCR kit
  • Pick up more candidates of amplicons to be tested using the Taq2X MPCR with set-up condition


Samples & Materials

  • Primers:
    • Blue primers mix: Y-1(122), 21-1(200), 1-1(270), 5-1(413), 22-1(757)
    • Yello primers mix: 11-1(131), 10-1(214), 15-1(297), 17-1(419), 16-1(806)
  • Templates
    • For all Qiagen MPCR (QMPCR) tests: 5 uL of 1 ng/uL human gDNA
    • For TaqPol2X MPCR (TMPCR) tests: 1 uL of Amplicon candidates picked up from human MDA (03/10/09): B10, E9, E10, F9, F10, H8, H9, H10


Procedures

  • QMPCR - no Q-solution
                            1 rxn      
    -----------------------------
    RNAse-free H2O           15.0 
    Template (1 ng/uL)        5.0   
    Qiagen kit enzyme (2X)   25.0 
    -----------------------------
                             45.0 (uL) 
    Add 5 uL primer Bule / Yellow
  • QMPCR - with Q-solution
                            1 rxn 
    -----------------------------
    RNAse-free H2O           10.0 
    Q-solution                5.0
    Template (1 ng/uL)        5.0   
    Qiagen kit enzyme (2X)   25.0 
    -----------------------------
                             45.0 (uL) 
    Add 5 uL primer Bule / Yellow
  • TMPCR
                            1 rxn       
    -----------------------------
    RNAse-free H2O           19.0 
    Template (1 ng/uL)        1.0   
    NEB Taq pol.(2X)         25.0 
    -----------------------------
                             45.0 (uL)
    Add 5 uL primer Bule