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الكميات المطلوبة لتحضير 50 ميكروليتر الحجم الكلي للتفاعل هي : Components Stock concentration Final concentration Template125 ng/μL7.5 ng/μL Primer F200.

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Presentation on theme: "الكميات المطلوبة لتحضير 50 ميكروليتر الحجم الكلي للتفاعل هي : Components Stock concentration Final concentration Template125 ng/μL7.5 ng/μL Primer F200."— Presentation transcript:

1 الكميات المطلوبة لتحضير 50 ميكروليتر الحجم الكلي للتفاعل هي : Components Stock concentration Final concentration Template125 ng/μL7.5 ng/μL Primer F200 ng/μL4.0 ng/μL Primer R200 ng/μL4.0 ng/μL dNTP mix2.0 mM each0.2 mM PCR buffer10 X1 X MgCl225 mM1.5 mM Taq polymerase5 U/μL0.05 U/μL H2O

2 الكميات المطلوبة لتحضير 50 ميكروليتر الحجم الكلي للتفاعل هي : Components Stock concentration Final concentration Volume μl Template125 ng/μL7.5 ng/μL3 Primer F200 ng/μL4.0 ng/μL1 Primer R200 ng/μL4.0 ng/μL1 dNTP mix2.0 mM each0.2 mM5 PCR buffer10 X1 X5 MgCl225 mM1.5 mM3 Taq polymerase5 U/μL0.05 U/μL0.5 H2O31.5

3 ComponentsStock concentrationFinal concentration Template125 ng/μL7.5 ng/μL Primer F200 ng/μL4.0 ng/μL Primer R200 ng/μL4.0 ng/μL dNTP mix2.0 mM each200 μM PCR buffer10 X1 X MgCl225 mM1500 μM Taq polymerase 5 U/μL0.05 U/μL H2O الكميات المطلوبة لتحضير 20 ميكروليتر الحجم الكلي للتفاعل هي :

4 ComponentsStock concentrationFinal concentrationVolume (μL) Template125 ng/μL7.5 ng/μL1.2 Primer F200 ng/μL4.0 ng/μL0.4 Primer R200 ng/μL4.0 ng/μL0.4 dNTP mix2.0 mM each200 μM2 PCR buffer10 X1 X2 MgCl225 mM1500 μM1.2 Taq polymerase 5 U/μL0.05 U/μL0.2 H2O12.6 الكميات المطلوبة لتحضير 20 ميكروليتر الحجم الكلي للتفاعل هي :

5 ما هو الحجم المطلوب إضافته إلى برايمر وزنه 23 نانو مول ليكون تركيزه النهائي 100 ميكرو مولر

6 1mol/L==>1M 1umol/ul==>1M 1nmol/ul==>1mM 1nmol/ul==>1000uM==>V(ul)=(Mass/Concentration)*1000 1nmol/10ul==>100uMV(ul)=(23/100)*1000 23nmol/230ul==>100uMV(ul)=230 ما هو الحجم المطلوب إضافته إلى برايمر وزنه 23 نانو مول ليكون تركيزه النهائي 100 ميكرو مولر

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