Part D: Lecture 34

Question 1

Subcloning (def.)

Answer 1

moving an insert from one vector to another

Question 2

plasmid vector size

Answer 2

1-3 kb

Question 3

insert DNA for plasmid vector size

Answer 3

< 15 kb

Question 4

BAC vectors stands for ___

Answer 4

Bacterial artificial chromosomes vectors

Question 5

BAC vectors size

Answer 5

100 kb E.coli fertility plasmid (sex pilus)

Question 6

insert DNA for BAC vector size

Answer 6

100 - 300 kb

Question 7

why make BAC clones?

Answer 7

clone all DNA from an organisms

Question 8

Making BAC clones procedure

Answer 8

1) isolate genomic DNA
2) shear into large random pieces (100 -200 Kb)
3) ligate into vectors
4) Transform E.coli cells

Question 9

method of shearing DNA into pieces for insertion

Answer 9

sonication (vibrations)

Question 10

genomic DNA library (def.)

Answer 10

collection of the total genomic DNA from a single organism stored in different vectors

Question 11

sonication produces

Answer 11

equal, uniform fragments

Question 12

RE isn’t used for shearing DNA because ___

Answer 12

DNA fragments would be different lengths

Question 13

cDNA is ___

Answer 13

complementary DNA (from mRNA)

Question 14

purpose of making cDNA is to __

Answer 14

clone all mRNAs from an organism

Question 15

Making cDNA procedure

Answer 15

1)isolate mRNAS with poly A tails
2)reverse transpiratase to DNA from RNA
3)degrade mrna
4) dna poly copies ssdna to dsdna –> cDNA
5)ligate into vectors
6) transform cells

Question 16

cDNA library (def.)

Answer 16

collection of the total genomic mRNA from a single organism stored in different vectors

Question 17

cdna of mrna that had introns in DNA

Answer 17

cDNA would contain only the exons

Question 18

in making cDNA libary, poly A tails are used to ___

Answer 18

isolate mrna

Question 19

cDNA library would only contain genes that ____

Answer 19

were active and made mRNA

Question 20

After BAC cloning/cDNA cloning, ____

Answer 20

DNA is sequenced before being stored at -70C (indexing the library-we know which DNA is in which clone)

Question 21

BAC clones are used for ____

Answer 21

FISH

Question 22

cDNAs clones are used for _____

Answer 22

GMOs