SM01 Mini3 Flashcards Preview

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Flashcards in SM01 Mini3 Deck (25):
1

restriction enzyme

aka restriction endonuclease

bacterial enzyme that cuts double stranded DNA at palindromal sites

 

2

bacterial role for restriction enzymes

"immune system"

enzymes will cut DNA/RNA of bacteriophages (viruses that infect bateria)

3

recognition sequence

4-8bp of double stranded DNA that make a palindrome

4

sticky ends

single stranded over hang after restriction enzyme has cut double stranded DNA

easier to ligate

5

blunt ends

no free nucleotides on either end after restriction enzyme has cut double stranded DNA

very hard to ligate

6

rDNA

recombinant DNA

engineered piece of DNA

restriction enzymes used to cut two different pieces of double stranded DNA that are then ligated together

7

replication vector

helps to amplify inserted gene or transgene in the host cell

ex. pBR322 & pUC19

8

expression vector

expression of cloned transgene in host cell

ex. anti-IL-23-Fc protein

9

cDNA

complementary DNA

created by reverse transcription of RNA

library lacks introns compared to genomic DNA library

10

hybridization probe

single or double stranded fragment of DNA/RNA (100-1000bp) tagged w/radioactive/fluorescent/enzyme marker to detect complementary target DNA presence in compex or gene library

may be chemically synthesized or cloned or via PCR amplification

11

allele specific oligoneucleotide

ASO

15-20bp

detects differences on nucleotide sequences between 2 alleles

helps detect gene mutations

12

southern blot

for DNA

13

Northern blot

for RNA

14

Western blot

for protein

15

restriction fragment length polymorphism

old technique used for detection of variation in DNA sequences via band profile comparison

disadvantages: need large amt of DNA, few loci detected, automation not possible→ time consuming & expensive

16

LacZ

reporter gene encoding for beta-galactosidase

found in pUC19 

recognition sites are in gene

if insert isn't present, colony is blue b/c beta-galactosidase is still active

if insert is present, colony is white b/c beta-galactosidase gene was cut & is inactive

17

agarose gel

separates DNA/RNA according to size

18

polyacrylamide gel

separates proteins or very small nucleic acids according to size & net charge

19

SNP

single nucleotide polymorphism

difference in DNA sequence amongst individuals/groups/populations

>1% of pop

causes: silent/harmless/latent/harmful effects

20

variable number of tandem repeats

short DNA sequence (40-100bp) repeated in a tandem fashion at specific chromosomal loci

varies between individual, but heritable- should possess one from each parent

used for DNA fingerprinting→forensics

<5bp= microsatellite (simple sequence)

minisatellite= >5bp, GC rich

21

steps of PCR

denaturation

annealing

elongation

termination

22

how to determine appropriate PCR primers

very specific & lack 2º binding site

16-24 nucleotides long

50% GC content

meting temp= 55-65ºC

23

ddNTP

chain terminators

3'OH is changed to H and thus cannot form a bond to teh next nucleotide (PO4 group)

 

24

oculocutaneous albinism

red eyes, bleach white skin & hair

autosomal recessive

25

linked markers

close together on same chromosome

optimal if very close together so they are not separated by recombination

used to mark a gene to predict carrier status if gene can not be directly detected or don't know the sequence of the gene