Flashcards in Devo Lect 3 - techniques Deck (20):
1
Gene cloning
Copying genes for study, from cDNA library or genomic
2
Promoter region
RNA poly and TF bind there, control on/off genes, KEY for developmental bio
3
DNA hybridization
complementary binding; southern blots; visualize presence of genes (probes); look for conserved across organisms
4
DNA sequencing
dideoxy nucleotides: fluoresce and terminate
5
northern blot
temporal and spatial (when and where) gene expression patterns; gel, transfer, probe
6
in situ hybridization
temporal and spatial (when and where) gene expression patterns; probe put on tissues, bind to complimentary DNA
7
PCR
amplifies segment of DNA; 95, 37, 72 - denature, anneal, taq polymerase
8
RTPCR
Real time or reverse transcriptase PCR - makes DNA from RNA, then PCR - shows genes that are even barely expressed (sensitive)
9
QPCR
Quantifies the amount of DNA; primer fluoresces so we can measure it by that
10
Expressing a gene to study
in xenopus oocytes, tissue culture, bacteria, transgenic organism and study effects
11
Knock out
insert some DNA to recombine (homologous sequence), get heterozygote, then breed to get homozygous knockout and study effects
12
Antisense DNA
insert little bits of complementary sequences which bind to mRNA, can’t be translated; not used that much
13
RNAi
use a very little bit of dsRNA, stimulates machinery of cell to cut up the RNA of choice
14
Western Blot
where proteins are expressed; need labeled antibody
15
immunofluoresence microscopy
fluorescent antibody so you can see proteins
16
Glofish
Transgenic zebrafish; designed as environmental detector; expresses GFP, RFP, etc. in muscles; insert muscle-specific promoter in front of GFP
17
Bioreactor purposes
A living system to grow large amounts of something, either to isolate something (ie. insulin, vitamins) or a useful protein (ie golden rice)
18
Glofish reproductive rate study
Slower reproduction; due possibly to colour; also due to energy expenditure to express GFP at high levels
19
Aquabounty atlantic salmon
Has chinook salmon growth hormone with promoter from Ocean pout - growth hormone is active all year rather than periodically in wild type
20