chapter 5 Exploring genes and genome Flashcards
southern blotting used for
DNA
restriction enzymes are
restriction endonucleases that cleave both strands of the recognised DNA
characteristic of cleavage sites when talking about RE
twofold rotational symmetry
when is polyacrylamide gel is used
what about more porous agarose gel
1000 bp
larger fragments as large as 20kbp
how to mmake sure that the chain termination in sanger sequencing will only take place occasionaly
by making the concentration of the deoxy analog very low
number of bp that can be sequenced using sanger sequencing
1000 bp
what is stringency in PCR how can it be controlled?
the required closeness of the match between the primer and target . by adjusting temperature and salt
when is the DNA only between primers amplified
at high temperatures
RFLPs
(restriction fragment length polymorphisms) polymorphisms within restriction sites that change the size of DNA produced using apropriate enzymes
what sources of energy does a ligase in RE use?
ATP or NAD+
how is the problem with not having restriction sites in DNA solved?
Solved by adding chemically synthesized DNA-linkers
what do plasmids carry?
antibiotics inactivating genes
toxin production
natural products degrading genes
polylinker region?
has many unique restriction sites within its sequence
Reporter genes
plasmids, easily detectable markers such as antibiotic resisting enzymes
expression vectors
plasmids for the production of large amounts of protein
Advantages of using modified vira
they enter bacteria much more easily than do plasmids
cosmids
hybrids of lambda phage and a plasmid used for the insertion of large DNA-inserts
what agent is used to denature DNA for DNA hybridisation
sodium hydroxide
which one is resistant to alkaline hydrolysis DNA or RNA
DNA
what enzyme is used for synthesizing the second strand when RNA-strand is hydrolysed in cDNA synthesis and what does it do
terminal transferase
adds G bases to the same strand
and then a primer of C-nucleotides is added to the newly synthesized complementary strand
The principle behind site-directed mutagenesis
using primers with one mismatch
inverse PCR why used and mechanism
used to for example introduce deletions, by using primers that orient in different directions to the ones used in normal
PCR
mobile genetic elements
non-coding DNA in the human genome, from retrovira inserted themselves over time.
example of mobile genetic elements in the non-coding DNA in the human genome
SINES (short interspersed elements)
LINES (long interspersed elements)
