biotechnology

Question 1

what’s PCR?

Answer 1

Polymerase Chain Reaction

• amplification of DNA
• used for Gel electrophoresis

Question 2

step 1 PCR

Answer 2

Denaturation

Separate DNA strands by breaking hydrogen bonds (94-96C)

Question 3

step 2 PCR

Answer 3

2. Annealing

Cool down to 50-70C

Question 4

STEp 3 PCR

Answer 4

3. Elongation

Taq polymerase builds complementary optimum (72C)

Question 5

how does gel electrophoresis work?

Answer 5

Wells always on positive side because DNA negatively charged

Phosphates (sugar-ribose backbone) migrate to the positive terminal

Question 6

what are ladders for?

Answer 6

isolate because it has known base pair lengths

Question 7

whats aragose gel for?

Answer 7

• Permeable
• Smallest fragments travel fastest
• Longer fragments travel slower (negative terminal)

Question 8

what’s sanger sequencing (- Must do PCR before sequencing)

Answer 8

method of sequencing: random of whether a deoxyribose or dideoribose is introduced to sequence

Question 9

what’s ddNTPs?

Answer 9

randomly inserted by DNA polymerase which terminates chain elongation