Mol Bio Exam 3 Final

Front 1

form of H bonds btwn ssRNA/DNA that are comple

Back 1

n acid hybridization

Front 2

btwn 2 polynucl chains which have compl bases
principle: at incr temp, dsDNA will denature > ssDNA, whn decr emp, strands anneal due to bp

Back 2

intermolecular hybridiz

Front 3

ssRNA/DNA oligonucl labeled with a reporter chem
will hypridize to target n.a. on the basis of bp

Back 3

probe

Front 4

DNA/RNA is immobilized on inert support so that self annealing is prevented
bound seq are available for hybrid with added probe
use filter paper (mc)

Back 4

solid support hybrid

Front 5

typer of solid support hybrid

Back 5

S blot
Dot/slot blot
N blot

Front 6

procedure for transf denatured DNA form an ag. gel to a solid support filter where it can be hybridized with a compl n.a. probe
DNA sep by size so that spec frags can be identified
DNA only

Back 6

S blot

Front 7

S blot commonly used to

Back 7

ident spec seq of DNA in which the frags are sep by electro> a memb and ident with probe
detects restric frags
to screen DNA and cDNA libraries

Front 8

to study gene expr with RNA
sep RNA Pattern > nylon memb
hybridize with 1 probe only and need separate blot for each probe tested

Back 8

N blot

Front 9

for multiple gene expr at once
thousands of probes on slide/memb
target is cDNA/mRNA
fluor labeled probe/target
monitors activity of genes by mRNA hybridiz

Back 9

Micro array

Front 10

DNA/RNA bound directly to solid filter
no size sep
for mult samples and quantity

Back 10

dot/slot blot

Front 11

id spec translocation breaks, chrom del, and gene amp
for solid tumor study

Back 11

FISH

Front 12

target n.a. is ampl

Back 12

PCR
NASBA
TMA
SDA

Front 13

signal is ampl

Back 13

HC2
bDNA

Front 14

probe is ample

Back 14

bDNA

Front 15

ampl DNA
thermal cycling
req: target, primers, Taq, buffer, dNTP

Back 15

PCR

Front 16

most sensitive for mRNA
starts with RT, mRNA> cDNA

Back 16

RT PCR

Front 17

simul ampl and quant target DNA
Taq man probe= reporter

Back 17

Real TIme PCR

Front 18

2 or more mRNA targets at once

Back 18

Multiplex PCR

Front 19

to determine order of nucl bases of DNA
Sanger method and maxam gilbert method

Back 19

DNA seq

Front 20

polymer chain is term with ddNTPs labeled with diff colors, detected by laser

Back 20

Sanger method
DNA seq

Front 21

detects spec proteins in sample
uses SDS PAGE to sep denatured proteins by length
uses 2ndary ab with color/chemil enzyme
for confirm of HIV, Mad cow, lyme disease

Back 21

W blot

Front 22

ideal candidates for blood genotyping

Back 22

Sickle cell
Thall
H anemia
lekemia
pst transf/ pos DAT
incr incidince ag for ID

Front 23

acceptable samples for genotyping

Back 23

EDTA blood
cheeck swab- babies and aplastic pts
leukored filters

Front 24

ideal donors for genotyping

Back 24

based on dempgraphic area
Blacks for sickle cell pts
screen for rares

Front 25

when to genotype

Back 25

prior to transf
to obtain extended pheno
post transf ab ID
DAT pos

Front 26

why genotyping is done in blood bank

Back 26

can incr donors by genot screening
incr care and TAT for pts
to find more accurate match
fro discrep and rare ab

Front 27

gold std for mol diag

Back 27

N.a. ampl

Front 28

use known n.a. probe labeled with detector to hybrib to another n.a. with bp
rapid, simple, cheap, accurate
req large number of orgs

Back 28

non ampl N.a. probes

Front 29

single/few copies of DNA/RNA are ampl by enzyme under direction of primer
rapid, sensitive, accurate
need extra sample prep, false pos

Back 29

target ampl

Front 30

target ampl methods

Back 30

PCR
NASBA
TMA
SDA
for N. gon, C. trach, M. tuber, HBV, HCV, HIV, CMV

Front 31

isothermal n.a. ampl based of restr enzyme and polym
for N. gon and C. trach
target1

Back 31

SDA

Front 32

signal ampl methods

Back 32

bDNA
HC2

Front 33

target n.a. seq not repl but ampl signal from probe
FDA abbroved for HIV and HCV

Back 33

bDNA

Front 34

signal ampl after hybrid of target DNA with RNA probe
incr sensitivity
FDA cleared for HPV

Back 34

HC2

Front 35

size of entire plasmid.chrom is determined
for genotyping or genetic fingerprint
gold std for patho orgs

Back 35

PFGE/ DNA fingerprint

Front 36

direct seq is determined, most accurate
for HepC

Back 36

N.a. seq

Front 37

Lim of N.a. ampl

Back 37

expensive
needed trained techs
contam is easy
rigid QC
need confirm test

Front 38

DNA is pos, org is absent
can be after antimicrobial therapy (C. trach and B. pert), gene present but not expressed (B. pert), gene present but unable to distiguish from normal flora ( S. pneumo)

Back 38

Clinical false pos, good

Front 39

from contam
cross reactivity

Back 39

Analytical false pos, bad

Front 40

can be caused by Norovirus, org rapidly cleared, DNA lost during processing, ampl inhibition

Back 40

False neg

Front 41

for detecting or correcting defects by estab limits of acceptable performance

Back 41

QC

Front 42

for continuing monitoring and improving the reliability, efficiency, and clinical value of lab tests
to decr and prevent errors
CLIA 88, CLSI, ACMG, CAP

Back 42

QA

Front 43

QA can be done

Back 43

in pre analyt, analyt, and post analyt of specimen processing

Front 44

to prevent contam/carry over

Back 44

good lab practice
sep pre and pst ampl
PPE
one way work flow

Front 45

poor collection or wrong source

Back 45

False neg, good

Front 46

poor PCR design, inhib PCR, PCR error

Back 46

False neg, bad

Front 47

techs for Fragile X

Back 47

RFLP
S. blot
PCR with RE
methylation

Front 48

techs for CF

Back 48

targeted/scanning mutation analysis with PCR
panel sensitivity

Front 49

CF

Back 49

CFTR mutation, F508 del is m.c.

Front 50

Fragile X

Back 50

trinucl repeat expansions of CGG on FMR-1 gene of X
mc form of inherited mental retardation
more Males

Front 51

Polymorphism

Back 51

germ line changes
in 1% of popl
RFLP
VNTR
STR
SNP

Front 52

mini satellite repeats
for DNA fingerprinting

Back 52

VNTR

Front 53

micro satellite repeats of CA, highly suscep to mutation when DNA repair mech lost
use PCR and tumor testing

Back 53

STR

Front 54

single nucl variations
detect with PCR or DNA Chip

Back 54

SNP

Front 55

for BRAF/KRAS

Back 55

sequencing, RT-PCR, hybridiz (Dot blot)

Front 56

for HER2

Back 56

FISH

Front 57

for HPV

Back 57

HC2, cervical swab, PCR/tumor spec

Front 58

causes 67% of melanoma
v600E= 90% of mutations
caused by GAG mutation

Back 58

BRAF

Front 59

mutations prevent GTP hydrolysis= always on > incr repl
most mutations in codons 12,13,61 on chrom 12
G>T = Val instead of Gly > abn growth

Back 59

KRAS

Front 60

predisp of BRAC1, 2 genes
predict with HER2 ampl target
FISH- fluor prob on tissue

Back 60

breast cancer

Front 61

all contain HPV DNA
HPV 16, 18, 31= 95%
use HC2, signal ampl
detects but doesnt distinguish

Back 61

Cervical cancer