RNA overview

Question 1

What are the differences between RNA and DNA sugars?

Answer 1

RNA sugars have two OH’s

Question 2

How does the extra OH group on RNA sugars effect the nucleotides?

Answer 2

make them less stable

Question 3

Whats the difference between RNA and DNA bases?

Answer 3

RNA uses U instead of T (u is less stable)

Question 4

RNA is synthesizes in which direction?

Answer 4

5’ to 3’

Question 5

What performs transcription? How?

Answer 5

rna polymerase does, reads template dna in 3” to 5” direction and add nuceltides in 5” to 3” directions

Question 6

RNA polymerase unwinds how many bp of dna at a time?

Answer 6

17

Question 7

About how much of the human genome is protein coding?

Answer 7

1-2%

Question 8

What does UTR stand for?

Answer 8

untranslated region

Question 9

What does RBS stand for?

Answer 9

ribosome binding site

Question 10

What are regulatory sequences in DNA? Do they get transcribed?

Answer 10

are regions that determine where and when the protein coding region will be transcribed (like in what tissue and what time), no they don’t get transcribed

Question 11

Does only the protein coding region get transcribed?

Answer 11

No, UTRs and introns, and RBS and terminators get transcribed

Question 12

Do prokaryotes have introns?

Answer 12

no

Question 13

What is an exon?

Answer 13

is included in mature RNA and translated to proteins

Question 14

What happens to introns during translation?

Answer 14

get spliced out

Question 15

What is the splicing machinery?

Answer 15

the spliceosome

Question 16

How does the spliceosome work?

Answer 16

It recognizes specific nucleotide sequences at the junction of exons and introns

Question 17

Introns begin with what and end with what?

Answer 17

Begin with GU and end in with AG

Question 18

What is the +1 site in dna?

Answer 18

Where transcription starts, rna polymerase recognizes the promoter here

Question 19

Whats the -1, -2, -3 site? what about the 1,2,3 site?

Answer 19

Is the diff position upstream after the promoter. Positive values are downstream of the promoter

Question 20

What are promoters?

Answer 20

They position RNA polymerase for transcription

Question 21

What is the sense strand?

Answer 21

Is the 5” to 3” strand, is also referred to as the coding strand

Question 22

What is the antisense strand?

Answer 22

Is the noncoding strand 3’ to 5’, promoter isnt here, also called the template strand where transcriptions builld ofof of

Question 23

What is the promoter sequences in all E.coli genes importance?

Answer 23

Sequences in the promoter (-35 and -10 regions) position the RNA polymerase for initiation of
transcription in E. coli

Question 24

What is quantitative PCR?

Answer 24

A method of PCR that allows us to quantify the amount of
DNA in the original sample

Question 25

What is the gold standard for covid 19 testing?

Answer 25

qPCR

Question 26

Does fluorescence increase as dna gets amplified?

Answer 26

Yes, after every cycle

Question 27

What is a CT value?

Answer 27

Cycle number at which the fluorescence level crosses the threshold

Question 28

Say one sample crossed threshold at CT 2 and the other did at Ct 34, which one has more dna in the original sample?

Answer 28

the staple that crossed the threshold at CT 2

Question 29

How do we use qPCR to monitor gene expression?

Answer 29

We convert mRNA to cDNA and then do pcr and incorporate dye.

Question 30

What is cDNA? How do you make it?

Answer 30

Is complimentary DNA, made by using enzyme reverse transcriptase on mrna

Question 31

You get a higher Ct value for one gene using cDNA, what does that mean?

Answer 31

That gene has higher expression (is transcribed more)