Sam:LabNotes/Microbiome/2009-3-13: Difference between revisions
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>Sam Chiang |
>Sam Chiang |
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Total= 14 rxns | Total= 14 rxns | ||
*Master solution of 20 uL QMPCR with Q-sol. | *Master solution of 20 uL QMPCR with Q-sol. | ||
Revision as of 07:51, 14 March 2009
Qiagen MPCR test with Q-solution on MDA L4N6/Tre amplicons
Objectives
- Test of 20 uL volumn of QMPCR with Q-sol
- Confirm the amplicons from MDA L4N6/Tre
Samples & Materials
- Primers: Note: use the un-adjusted primer mixture
- Blue primers mix: Y-1(122), 21-1(200), 1-1(270), 5-1(413), 22-1(757)
- Templates: (Amplicon label form MDA reaction)
Row A: B1 B2 B3 B4 B5 B7 B8 B: C1 C2 C3 C8 A4 gDNA H2O Note: A4 = p-pos Total= 14 rxns
- Master solution of 20 uL QMPCR with Q-sol.
1 rxn 16 rxn ------------------------------------------------- RNAse-free H2O 5.0 80.0 Q-solution 2.0 32.0 Primer mix (Blue) 2.0 32.0 Qiagen kit enzyme (2X) 10.0 160.0 ------------------------------------------------- 19.0 304.0 (uL) Add 1 uL template
- QMPCR programs: QM-PCR52
Step1 95C, 15m Step2 94C, 30s Step3 52C, 1m30s Step4 72C, 1m30s Step5 Goto Step2 X 30 Step6 72C, 10m Step7 4C, forever
Results
File:ZhangLab 2 2009-03-13 10hr 09min-with annotate.bmp
- Gel Electrophoresis
- 2 % agarose: (1.5 g agarose +100 mL 0.5X TBE + 8 uL SYBR safe) -> two medium tray with 13-well comb
- Loading: Sample 10 uL + 2 uL loading buffer; Diluted ladder 10 uL
- Run at 135 V for 30 min.