When strands of DNA or RNA are created in a cell (or in PCR), are new nucleotides added to the 5’-end or 3’-end?

When strands of DNA or RNA are created in a cell (or in PCR), are new nucleotides added to the 5’-end or 3’-end?
What are the three parts of a nucleotide?
How many hydrogen bonds do the base pairs of adenine and thymine have? How many do cysteine and guanine have?
In the plasmid isolation lab, where/what are we isolating plasmids from?
What is a plasmid?
Double stranded piece of linear DNA that copies itself in a cell.
Double stranded piece of circular DNA that stays separate from the cell’s genomic DNA and copies itself in the cell.
Piece of DNA that integrates/joins into the cell’s genomic DNA.
Double stranded RNA that denatures to give two mRNA transcripts.
Our isolated plasmids were analyzed by loading them into an agarose gel and then separating them based on size by applying an electrical current to the gel. What does an intact, circular plasmid look like when it is run out on a gel (what is the typical banding pattern)?
What controls were set up in reverse transcription?
Why are we using reverse transcription to get our actin sequence, and not just using PCR?
What are the three steps in a PCR cycle?
Don’t miss the bonus question below!
Bonus question (1 point):
In class we discussed how our actin primers were designed. For practice, design a set of primers that will amplify the coding sequence for Dsg3. Using the sequence found here:
https://www.ncbi.nlm.nih.gov/nuccore/NM_001944.3
(Links to an external site.)
first determine where the coding sequence is. (HINT: if you scroll down on the page, you’ll come to “CDS”. Click on it and it will highlight the entire coding sequence of Dsg3. This is the sequence you want to copy through PCR).
Once you see where the coding sequence is, design your forward and reverse primers. For your primers,
Both primers should be 18-24 bases long (ours in lab are usually 20-21).
The forward primer should include or be before the start codon (the “ATG” that starts at 85), and will be the same sequence as the mRNA sequence shown for Dsg3.
The reverse primer should include or be located after the stop codon. For Dsg3, the stop codon is “TGA”. Note that the reverse primer sequence needs to be the reverse and complement of the sequence shown in this entry.

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