PLEASE, I NEED TO DESCRIBE EVERY STEPS REALLY WELL. STEPS TO HANDLE AND PREPARE THE SAMPLE...

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Biology

PLEASE, I NEED TO DESCRIBE EVERY STEPS REALLY WELL. STEPS TOHANDLE AND PREPARE THE SAMPLE AND THEN, STEPS OF RNAseq.
Sequencing RNA from a single circulating tumour cell:
A research laboratory has isolated a CTC from a patient andwould like a description of how you would process the sample andhow you would sequence all nonâ€coding RNA from the cell.
a) Describe the steps involved in HOW you would prepare the RNAfor RNAseq;
b) HOW RNAseq works.
Also explain WHY each step was done and consider the mostappropriate method for isolating nonâ€coding RNAand removing rRNA from your sample.

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a ISOLATION OF RNA FOR RNA seq The three major techniques extensively used for RNA extraction organic extraction such as phenolGuanidine Isothiocyanate GITCbased solutions silicamembrane based spin column technology and paramagnetic particle technology One of the most commonly used methods is the phenolGITCbased organic extraction But RNA samples isolated by this method are frequently contaminated with proteins organic solvents such as phenolchloroform salts and ethanol SIlica membrane based spin column is used in laboratories for isolation of RNA from patient samples Isolation kits like Qiagen are readily available for RNA isolation and the purity is also high HOMOGENIZATION OF THE TISUUE 1 TRIzol reagent in a 50 ml at room temperature RT before taking the frozen See Answer

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