13. hat is a phenol/chloroform extraction (see lab manual)? When we used this extraction in the RNA isolation protocol.

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13 Hat Is A Phenol Chloroform Extraction See Lab Manual When We Used This Extraction In The Rna Isolation Protocol 1 (80.49 KiB) Viewed 51 times

13. hat is a phenol/chloroform extraction (see lab manual)? When we used this extraction in the RNA isolation protocol. What was in each layer and what layer did we discard/keep? 14. On PCR: a. What are some guidelines for primer design? (Tm, GC%) Why do we have these guidelines in the context of the PCR reaction itself? b. What is different between PCR primers and codons"? c. What is ‘hybridization' in the context of primers? 15. What do we quantify nucleic acids so often? What could adding too much of a nucleic acid do to a PCR reaction? Too little? 16. How does gel-electrophoresis work? What charge is DNA and how do gels separate DNA? What determines the size of DNA? 17. What is sense vs anti-sense in relation to mRNA? What is the relationship of sense to our WISH probe preparation? 18. When sequencing, we use these special nucleotides to terminate the reaction while tagging the last nucleotide. 19. What role do the T7 and SP6 promoters play in our WISH probe preparation? What reaction are they specifically involved in? 20. Extra Credit: Calculate PCR Primer solution preparation recipe. Examples for practice a. 0.2uM when adding 2ul of primer with 25nmole provided by vendor b. 0.25uM when adding lul of primer with 25nmole provided by vendor