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Core I > exam 3 > Flashcards

exam 3 Flashcards

1
Q

four main parts of mass spec

A

separation source front end
ionization like MALDI
mass separater
detector

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2
Q

dideoxy sequencing

A 
3' Oh group is gone
stop short
modern diff color per end
or run each column differently
modern is color capillary
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3
Q

Pacbio

A

single long strand
stuck in well with poly adding one at at time
suuuuper long DNA

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4
Q

ilumina

A

blob with primers

added one at a time to get colors

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5
Q

RNAseq

A

make cDNA with reverse transcriptase
use something like illumina
know what relative levels are active
super expensive though

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6
Q

microarray

A 
littlespots
use cDNA again
has oligonucleotides single stranded
bind complimentary
to probe for a bunch of genes and concentrations 
intercalated dies
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7
Q

HPLC

A

high pressure liquid chromatography
samples are run ofer a column w/ high pressure columns containg resins like (affinity, pH, size exclusiong) to attract specific samples samples bound to resin can be taken in a specific order by washing organic solven in increaseing concentrations over the column

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8
Q

PCR

A

polymerase chain reaction
melt - 94
anneal primers - 50-65
elongate/amplify -70s is when taq polymerase is most active

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9
Q

Tm

A

50% of primer is bound

hotter than that less primer is bound

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10
Q

where when pluripotent mouse ESC

A

inner cell mass of a 3.5 day old mouse blastocyte

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11
Q

yeast 2 hybrid model

A

for proteins
half of an enhancer on each of 2 proteins
if they interact they activate enhancer to turn on promoter on a reporter gene

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12
Q

Northern Blots

A

RNA
run through a gel
hybridize to probe
peel and loook

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13
Q

Southern Blots

A

DNA

same steps

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14
Q

Western Blots

A

Protein
denature first
antibodies to do bait and hook

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15
Q

CRISPR-Cas

A

target specific gene with CAS9 and guide protein in 0.5 fertilized egg (pronucleus) then transfered to pseudopregnant female mouse

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16
Q

RNA absourbance

A

260/280

about 2

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17
Q

RNA absorbance

A

260/280

about 2

18
Q

DNA absirbance

A

260/280

about 1.8

19
Q

ligase independent

A

how do they associate

20
Q

ligase independent

A

use false sticky ends instead

21
Q

genome library

A 
restriction enzyme to randomly cut
cut vector the same
transform bacteria 
grow
harvest
see if hybridize with probe
22
Q

cDNA library

A

put it in plasmid
up-regulate
find exons and introns
cut and see if DNA binds to it

23
Q

CRISPR

A 
delete need two
add need two and donor plasmid
insertion need one and donor 
in the one cell stage 
into zygote 
gRNA can be bought if we know sequence 
makes humans homo
24
Q

ZFN and TALENS

A

same as crisper but find it with their modules

25
Q

RNA interference

A

in something feed words
putting in si and miRNA
because double stranded RNA is unnatural
can be incorporated

26
Q

Cre-lox

A

intersect of reporters for cre and the gene
tissue specific
can be in adults not just embryos
can knock out things later that would be fatal to embryos

27
Q

2d electrophoresis

A

for proteins
step one by PI
second By SDS to do by size mass ratio s
stain after blue

28
Q

stain first

A

DNA gels ethidium bromide

29
Q

pull down

A

tag with antibodies
size
similar to affinity chromatography

30
Q

sucrose gradient centrifugation

A

dense on bottome
settle in and then collect bottom layer first
S units

31
Q

FPLC

A

doesnt denature

size exclusion

32
Q

reverse phase HPLC

A

hydrophilicity
phobic column
polar solvent
normally phases are opposite

33
Q

MALDI

A

laser bounces off dish

34
Q

DESI

A

spray evaporate hit

35
Q

Quadripole

A

narrow hole

36
Q

tof

A

mirrors

longer bigger

37
Q

orbitrap

A

round a magnet

longer bigger

38
Q

SRM MRM

A

single and multiple reaction monitoring

single vs multiple proteins during like MS

39
Q

MudPIT

A

multiple dimensional Protein ID Tech
proteins to poly peptides compare to known
bottom up

40
Q

iTraq vs silac

A

isobaric attatch known size to protein

silac is heavy light from tagging with 15N

41
Q

imaging MS

A

MS with pic instead of electron multiplier tells you location
slice not cell sized
kinda like a heat map

42
Q

gene targeting in embryonic stem cells

A

long arms in yeast

cut elsehwer

Core I (6 decks)

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