rrna genes-quiz 4

Question 1

bacterial 16s ribosomal subunits

Answer 1

• have conserved regions of similarity. this provides universal locations for pcr primers.
• have divergent regions of dissimilarity. this provide sequence variability used to identify taxa.

Question 2

pcr

Answer 2

dsDNA denatured into single stranded dna. 95. 2 min. 15 sec.
two primers bind to opposite strand to prime in opp directions. 54. 15 sec
taq polymerase replicates template dna using single stranded dna primer. 72. 8.5 min.

Question 3

pcr is an enzymatic reaction. requires
buffer
enzyme
enzyme cofactors

Answer 3

buffer solution, dNTP, whole bacteria
enzyme cofactors: Mg++
enzyme- taq (thermus aquaticus)

Question 4

why use Mg++?

Answer 4

dna ends are negative bc of phospholipids, so in order to bridge them, use mg+. - and + come together.

Question 5

how is dna sequence determined?

Answer 5

using ddNTP Sanger sequencing. primary data is electropherogram. (can include undetermined bases “N”’s, usually at beginning and end.

Question 6

pcr undergoes

Answer 6

30 cycles

Question 7

electrophoresis has

Answer 7

agarose gel