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Comparison of Different Methods of mRNA Quantitation


FeatureNuclease Protection AssaysNorthern BlottingRT-PCRIn Situ HybridizationRelative and Absolute Quantitation
Yes
Yes
Yes
Yes
Detection Limit (Copies of mRNA)
4,000-5,000
10,000
1 (theoretical)
Medium
Level of Optimization Required
Medium
Medium
High
High
Sample Size
100 g max
30 g max
Not critical
Tissue sections
Detect Multiple Transcripts
Yes, in same tube
Yes, ge
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Calcium Phosphate Transfection Kit from Invitrogen
Custom siRNA, HPLC purified from Ambion
Silencer Pre-designed siRNA, standard purity, annealed from Ambion
Minimal Essential Medium Alpha Medium (Custom) from Invitrogen
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