MCDB 423 Lecture Notes - Lecture 13: Ribonuclease, Agarose Gel Electrophoresis, Pipette
23 Dec 2020
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Step 2: in vitro transcription: reaction components. Assemble the reaction at room temp: spermidine in the cold 10x bu er will cause dna to precipitation. Step 3: destruction of template dna: use dnase1 to degrade template dna after the rna is synthesized, get rid of dna keep rna. Licl does not precipitate dna proteins or carbohydrates: after you add you want to mix thoroughly and chill overnight at -80 degrees c. Precautions while working with rna: rna degreased very easily and rnases are everywhere, rnase-free solutions, depc (diethyl-pyrocarbonate) destroys rnases, bench and work area rnase-free, rnase-free material like pipette tups, micro-centrifuge tubes, etc, always wear gloves. Hands are excellent source of rnases: keep rnase-free tubes and pipette boxes close as much as possible, prevent taking at your samples. In situ hybridization: northern blots (rna blots) Signals seeing on in situ hypnotization are signals that can be detected by rna probes.
