NEB CUTTER

Question 1

1. How would you describe the extension produced by Ac1l in terms of its length and whether
   it is a 5’ extension or a 3’ extension.
   a. 5’: Two Nucleotide Extension
   b. 3’: Three Nucleotide Extension
   c. 3’: Two Nucleotide Extension
   d. 5’: One nucleotide Extension

Answer 1

a. 5’: Two Nucleotide Extension ✓

Question 2

Go to NEBCutter page and click on list of well known pieces of DNA. Choose pBR322 and
click submit button. Cut pBR322 with the enzyme Ac1l. How many fragments are generated?
a. 4
b. 5
c. 6
d. 8

Answer 2

4

Question 3

Write down the names of the first enzyme on the one cutter list and the last enzyme on the
one cutter list.
a. Acal and XmvII
b. BanHI and YpsIV
c. AatII and ZeaI
d. AIqV and Zral

Answer 3

c. AatII and ZeaI ✓

Question 4

Cut lambda DNA using the enzyme AF1II. Use the zoom function to zoom in to the region
surrounding the middle of the three AF1II cut sites. You will need to zoom in several times to
see the actual letters of the sequence in that region. Now. Looking at the top strand only,
write down the sequence of the ten nucleotides that lie upstream of the AF1LL palindromic
recognition sequence.
a. 5’ – TGAAATTCTC – 3’
b. 5’ – CTAAATTCTG – 3’
c. 5’ – AGATATGCTG – 3’
d. 5’ – AGAAATTCTG – 3’

Answer 4

d. 5’ – AGAAATTCTG – 3’

Question 5

Go to NEBCutter and click on the list of well known pieces of DNA. Choose pBR322 and click
submit. What NEB buffer is recommended to be used when cutting DNA with AF1III?
a. 1.1
b. 2.1
c. 3.1
e. I don’t know

Answer 5

3.1

Question 6

How many nucleotides are there in the extension produced by BamHI?
a. 4
b. 5
c. 6
d. 3

Answer 6

4

Question 7

Cut lambda DNA with two enzymes together – this is known as a double digest. Use the
enzymes AvrII and BamHI. How many fragments are generated.
a. 17
b. 12
c. 5
d. 8

Answer 7

8

Question 8

What is the size of the second largest fragment produced by cutting pBR322 with Ac1I?
a. 1384 bp
b. 1297 bb
c. 20044 bp
d. 976 bp

Answer 8

1297

Question 9

Go to NEBCutter and click on the list of well known pieces of DNA. Choose pBR322 and click
submit. What NEB buffer is recommended to be used with the restriction enzyme AhdI?
a. 1.1
b. 2.1
c. 3.1
d. CS
e. Don’t know

Answer 9

CS

Question 10

What is the size of the fifth largest fragment generated when lambda is double digested
using AvrI and BamHI?
a. 5505 bp
b. 4844 bp
c. 4515 bp
d. 3827 bp

Answer 10

5505

Question 11

s Ac1L digestion of pBR322 sensitive to methylation (GpG type of methylation)?
a. Usually
b. Rarely
c. Yes
d. No

Answer 11

yes

Question 12

What is the size of pBR322?
a. 4825 bp
b. 4361 bp
c. 5428 bp
d. 3893 bp

Answer 12

4361

Question 13

Go to NEBCutter and click on the list of well known pieces of DNA. Choose pBR322 and click
submit. pBR322 is what type of DNA?
a. Chromosomal
b. Circular
c. Linear
d. Viral

Answer 13

Circular

Question 14

Does BamHI have any neoschizomers or isoschizomers?
a. One neoschizomers and two isoschizomers
b. Yes
c. No
d. No neoschizomers but two isoschizomers

Answer 14

no

Question 15

When BamHI cuts across double-stranded DNA, it produces a
a. 3’ extension
b. 5’ extension
c. 5’ extension on one side and a blunt end on the other side
d. 5’ extension on one side and a 3’ extension on the other side

Answer 15

b. 5’ extension

Question 16

What is the accepted single-strand recognition sequence for BamHI?
a. GGATCC
b. CCTAGG
c. Both of the above
d. Neither of the above, the sequence must be written down in double- strand
format.

Answer 16

a. GGATCC ✓

Question 17

17. Looking at the screen press the correct button that will provide a list of enzymes that cut
    pBR322 just once. How many enzymes do cut pBR322 just once?
    a. 39
    b. 49
    c. 37
    d. 63

Answer 17

49