Biotech - PCR (DNA Sequencing, slides 1-10) Flashcards
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| 319064880 | because DNA encodes for all of the genes in an organism, the ___ of DNA is very important | sequence | |
| 319064881 | DNA is sequenced by essentially ____ DNA in the presence of special nucleotides which result in ____ of DNA | replicating, fragments | |
| 319064882 | sequencing technology relies upon the fact that ___ ___ needs a template | DNA polymerase | |
| 319064883 | sequencing technology relies upon the fact that DNA polymerase needs a ___ | template | |
| 319064884 | DNA is copied by reading the template strand and predicting the newly synthesized strand using __ __ rules | base pair | |
| 319064885 | T/F: DNA replication requires a template | true | |
| 319070828 | the key component in sequencing is the incorporation of a special nucleotide called a ____ nucleotide | dideoxy | |
| 319070829 | dideoxy nucleotides do not have a ____ so they are incapable of forming nucleic acids | 3'OH | |
| 319070830 | what has no 2'OH or 3'OH? | dideoxy nucleotide | |
| 319070831 | dideoxy nucleotides can only be added at the ___ (beginning/end) of a DNA strand | end | |
| 319070832 | dideoxy nucleotides (ddNTP) fall into that class of __ __ b/c once they are incorporated they halt synthesis | chain terminators | |
| 319070833 | if ddNTP is included, ___ of DNA are generated, each "capped" with a ddNTP | fragments | |
| 319070834 | what is the specific ddNTP determined by? | template strand | |
| 319070835 | what happens? ddNTP is added in place of ONE of the nucleotides (ex. ATP, CTP, ddGTP) | synthesis stops | |
| 319070836 | slides 4-6 show good representations of dideoxynucleotides | ... | |
| 319070837 | what happens? a small amount of ddNTP is added and ALL FOUR dNTPs are present? (ex. ATP, CTP, GTP, TTP, ddGTP) | two DNA fragments are produced (b/c either GTP or ddGTP can be incorporated) | |
| 319070838 | to determine how many DNA fragments there are you can use ____ | electrophoresis | |
| 319070839 | T/F: electrophoresis separates DNA fragments based on size | true | |
| 319074739 | smaller pieces of DNA will show up at the ____ (bottom/top) of the electrophoresis gel | bottom (b/c they can move farther) | |
| 319074740 | ____ (smaller/larger) pieces of DNA will show up at the top of the electrophoresis gel | larger | |
| 319074741 | if you have ATP, CTP, GTP, TTP, and a small amount of ddGTP, what will every fragment end in? what is the exception? | every fragment ends in G, exception is the full-length piece | |
| 319074742 | the size of the DNA fragment is based upon the number of ___ ___ | base pairs | |
| 319074743 | slide 8 is a good one for a visual of electrophoresis | ... | |
| 319074744 | to determine the sequence by looking at electrophoresis gel, you start at the ____ of the gel and work your way ___ | bottom, up | |
| 319074745 | the bottom of the electrophoresis gel represents the ___ end and the top of the gel is the ___ end | 5', 3' | |
| 319074746 | with electrophoresis, we are determining the sequence of the ___ strand | complement | |
| 319074747 | T/F: when using electrophoresis gel, the sequence of the template strand is determined | false (complement) | |
| 319074748 | T/F: when using electrophoresis gel, the sequences of the complement strand is determined | true | |
| 323719274 | does the primer show up on electrophoresis gel in DNA sequencing? | no (the gel shows the band that corresponds to the last nucleotide) | |
| 323719275 | T/F: the primer should be complementary to the sequence of template strand | true | |
| 323719276 | in DNA sequencing, the electrophoresis gel shows the _____ strand of DNA | complement | |
| 323719277 | at the bottom of the gel of DNA sequencing the first nucleotide that shows up is next to the ____ | primer |