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==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 mix Bule / Yellow (Use the unadjusted set -> equal ammount of each pair) *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 mix Bule / Yellow (Use the unadjusted set -> equal ammount of each pair) *TMPCR '''Note: only 1 uL template was used since MDA amplicon have have very high concentration 1 rxn ----------------------------- RNAse-free H2O 19.0 Primer Bule 5.0 (Use the djusted set) NEB Taq pol.(2X) 25.0 ----------------------------- 49.0 (uL) Add 1 uL MDA amplicon *QMPCR programs: QM-PCR52 '''Note: cycles were decreased from 35 to 30''' Step1 95C, 15m Step2 94C, 30s Step3 52C, 1m30s Step4 72C, 1m30s Step5 Goto Step2 X 30 Step6 72C, 10m Step7 4C, forever *QMPCR programs: TM-PCR52 '''Note: cycles were increaded from 35 to 40''' Step1 94C, 2m Step2 94C, 40s Step3 52C, 40s Step4 72C, 1ms Step5 Goto Step2 X 40 Step6 72C, 10m Step7 4C, forever
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