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 No edit summary |
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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 | ||
== | |||
==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