human genome and mutation detection part1/2

Question 1

how much nuclear DNA codes for functional gene products

Answer 1

2.5%

Question 2

where do you find polymorphic DNA sequences

Answer 2

in extragenic sequences

Question 3

what are polymorphic sequences closely linked to

Answer 3

linked to defined markers like gene, PCR primer. can also be linked to mutated genes.

Question 4

what are minisatellite sequences used for

Answer 4

to determine paternity and to map mutant genes and detect individuals

Question 5

what is another name for minsatellite sequences

Answer 5

VNTR variable number of tandem repeats

Question 6

how can you detect VNTRs

Answer 6

by multi locus probes detect different but related sequences.
single locus probes identify one specific sequence

Question 7

what does DNA profiling target

Answer 7

STR short tandem repeat

Question 8

how do you amplify STR sequence

Answer 8

by PCR

Question 9

how are PCR products analysed

Answer 9

by capillary electrophoresis

Question 10

what is LCN low copy number of amplification

Answer 10

it amplifies DNA from a very small cell numbers

Question 11

name a type of indirect DNA detection

Answer 11

by linkage analysis. You link polymorphic DNA markers with disease.

Question 12

what can direct detection do

Answer 12

can detect deletions, insertions by PCR or southern blotting. can detect point mutations.

Question 13

give examples of diseases caused by point mutations

Answer 13

sickle cell haemoglobin
alpha1-antitrypsin

Question 14

give examples of diseases caused by deletions

Answer 14

cystic fibrosis
muscular dystrophy
thalassaemia

Question 15

give examples of diseases caused by insertions

Answer 15

LDL receptor hypercholesterolaemia

Question 16

how does allele specific oligonucleotides work

Answer 16

you need to define hybridisation and washing conditions precisely. need to label the oligonucleotide.

Question 17

what is ARMS

Answer 17

amplification refractory mutation system

Question 18

name some detection systems

Answer 18

1)specific detection of amplified products-Taqman, molecular beacons
2)non specific - SYBRG in real time PCR
3) fluoresence transfer and chemiluminescences system
4) scorpion probes

Question 19

how can you detect known mutations

Answer 19

by allele-specific oligonucleotides hybridisation, PCR, gene chip

Question 20

how do you detect unknown mutations

Answer 20

by DNA sequencing

Question 21

what is first generation sequencing

Answer 21

shotgun sequencing- done by synthesis. High accuracy. Long read lengths. small data generated. example: ABI capillary sequencer

Question 22

what is second generation sequencing

Answer 22

massively parallel sequencing- by synthesis, less starting material needed, high accuracy, short read lenghts. example; MiSeq (illumina) Ion Torrent

Question 23

what is 3rd generation

Answer 23

single molecule sequencing- isngle molecule templates, low accuracy, long read lengths. example : single molecule real time sequencing. MinION