Ch 20 DNA Tools and Biotechnology

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1

DNA sequencing is what

genes complete nucleotide sequence

2

-DNA cloning

-gene cloning

identical copies

3

plasmids-

small dna molecules that replicate seperately

4

recombinant DNA molecule is what

piece of DNA inserted into plasmid. Now has DNA from two sources (plasmid and DNA from where inserted)

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cloning vector

plasmid used to clone a foreign gene

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bacterial plasmds common why?

amplify genes for research; readily available, multiply quickly

7

restriction enzymes

cut DNA at restriction sites, leave what is called restriction arrangements, one side is sticky side ; how bacteria enters plasmid

8

DNA ligase does what

seals bonds made through restriction fragments

9

Palandrome-

same bases on top and bottom;

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Sticky side wants to find complementary base, true or false?

true, wants to find compl

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gel electrophoesis

gel made of polymer seperates into currents negative and positive; separates based on size etc

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polymerase chain reaction- PCR process

heating (denaturization), adding primer, extnded with nucleotides

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Bacteria to eukaryotes

expression vector-cloning vector with highly active bacteria promoter- this overcomes differences

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difference bacteria to eukaryotes

difference in sequencing with promoters and other DNA sequences

2. presence of introns; use only cDNA which only has exons

yeasts sometimes used, cultures mammals and insects used

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hybridization

fluoresent dyes attached to probes to find mRNA

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RT PCR

compare amounts in several samples at same time

make cDNA which serves as template for amplification

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DNA microassays

finding networks; comparing thousands of genes at same time

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RNA sequencing

cut, converted to cDNA and sequenced

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how to determine gene function

mutagenesis-mute clones see what features missing

RNAi- break down or block genes mRNA

20

stem cells are

unspecialized but can be specialized under certain circumstances; reproduce infinitely

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topitotent

all potent cell

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dediffernciate

plants can go back and give rise to new specialized cell types

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nuclear transplantation

less differenciated/specialized more liekly it will work

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pluipotent

embryotic stem cells- capable of becoming differnt cell types

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main goal of cloning is what

to treat diseases

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induced pluripotent stem cell

use retroviruses to induce copies of 4 master regulatory genes. act like stem cells

27

gene therapy

introduction of genes for theraputic purposes. use of viral RNA and inject to become permanent

Bone marrow cell!!!