Analyzing the structure & function of genes Flashcards
DNA Cloning
Cloning a certain gene
_____________ cut DNA molecules at specific sites
Restriction enzymes
Restriction Enzymes(restriction nucleases)
Cut the DNA strand at a specific nucleotide sequence & prevent the transfer of DNA between strains of bacteria
____________ seperates DNA fragments that are left after the restriction enzymes cut the DNA strand (can be separted by size)
Gel electrophoresis
Once the ________ has been cut the fragments are loaded onto the gel on the negative charges side & when a voltage is applied to the gel the negative charged DNA strand moves towards the positive end of the gel & eventually the DNA fragments are spread out through the gel in groups based on sized (looks like a ladder)
DNA Strand
To ________ the certain fragments of DNA its removed from the gel by a scapel
Isolate
__________ begins with the production of recombinant DNA
DNA cloning
Recombinant DNA
Is a DNA molecule that contains a combination of nucleotide sequences from other individual DNA molecules (The restriction enzymes cut the two individuals DNA molecules at a certain nucleotide sequences and the DNA ligase join the two different DNA molecules to together to form the recombinant DNA molecules)
_____________ is the result of the two different DNA fragment joining together by DNA ligase gluing the fragments together
Recombinant DNA
Plasmid
Carry the DNA fragments that are supposed to be cloned (They are circular double-stranded DNA cloning vectors)
The _________ contains its own replication origin which allows it to replicate in bacterial cells where the certain recombinant fragments will be produced in large amounts
Plasmid
The ________ can also be cut open to allow the insertion of other DNA fragments (They are cut open by a restriction enzyme at a single site)
Plasmid
A fragment of the ___________ is attached / linked to the plasmids to be copied (Attached by the DNA ligase)
Recombinant DNA
A recombinant DNA can be copied inside a __________
Bacteria Cell
An entire genome can be represented in a ___________
DNA Library
DNA Library
Collection of cloned DNA fragment in bacteria
Genomic Library
A collecction of all the different genes that make the whole genome
cDNA Library
Contains the DNA copied from the mRNA
Complementary DNA (cDNA)
Is formed by the reverse transcriptase enzymes & the DNA polymerase where it is a complementary starnd to the mRNA
cDNA is made from _________
mRNA
__________ clones are made from fragment of the gene & also contains introns (noncoding DNA strands)
Genomic DNA
____________ clones are made from mRNA so they contain extrons that are used to produce proteins
cDNA
__________ provides a sensitive way to detect specific nucleotide sequences
Hybridization
DNA denturation
Process where the hydrogen bonds are broken & the DNA strands are seperated from one around (Done by heating DNA strand)
DNA renaturation (Hybridization)
Process where the the two seperate DNA strands join back together reforming their hydrogen bind base - pairs
___________ can be used for detecting any nucleotides sequences by using a probe that has complementary base-pairs to the wanted nucleotide sequences (for both DNA & RNA)
Hybridization
_________ & ________ libraries are used to clone large gene & need bacteria to copy genes
Genomic & cDNA
Polymerase chain reaction (PCR)
A better way of cloning genes & RNA & copied in test tubes
________ uses DNA polymerase & specific DNA primers to make (amplifying ) DNA sequences in a test tubes
PCR
For ______ to work it needs DNA hybridization to select certain nucleotide sequences & DNA ploymerase to copy that strand
PCR
The ___________ need a primer to start & the PCR also directs the polymerase to the right nucleotide sequence
DNA polymerase
_______ occurs in cycles where to start each cylce the two strands of the double - stranded DNA template are seperated & the primer is hybridized & the DNA polymerase then replicate the strand
PCR
_______ is used for cloning short DNA fragments or RNA which it can result in obtaining either a genomic clone or a cDNA clone of mRNA & doesnt need a library for cloning
PCR
The ________ dimers direct the cloning process of the certain DNA fragments in a test tube
PCR
______ can be used for diagnostic & forenstic application (can be used to detect infection, track epdiemics, forensic investigation, etc)
PCR
_________ can be used to detect the presence of a viral genome in a sample of blood
PCR
__________ depends on the analysis of DNA chains terminated at every position
Dideoxy sequencing
Dideoxy Sequencing (Sanger Sequencing)
Uses DNA polymerase along with dideoxyribonucleoside triphosphate
Dideoxyribonucleotide triphosphate
Are special chain terminating nucleotide & it make partial copies of the DNA fragment to be sequenced & produced a collection of different DNA copies that are terminated at every position in the orginal DNA sequence
The _________ method of sequencing DNA relies on chain terminating dideoxynucleotide triphosphate (ddNTPs)
Dideoxy
Automated ___________ relied on a set of four ddNTPs each bearing a uniquely flourescent tag
Dideoxy
