Sequencing DNA reactions?
“You set up four DNA sequencing reactions: (i) with terminator G* deoxyribonucleotides labeled with a red fluorescent dye; (ii) with terminator C* labeled with a green dye; (iii) with terminator T* labeled with a yellow dye; (iv) with terminator A* labeled with a blue dye. Each reaction also includes all four of the normal nucleotide precursors for DNA (A, G, T, C).
Each sequencing reaction includes a primer with the sequence
5’-TACG-3’
The sequence of the template strand you are sequencing is:
5’-CAACAAGCTCCAACCAAGAATAGCCCCGTA-3’
For the reaction with each terminator, write down the sequences of each of the newly made DNA strands that would be produced (each ending with a terminator)”
I don’t even know where to start. Any guidance would be appreciated, my book is useless and Google hasn’t been helpful either.
Well, do you know how to create the basic complementary strand? If so, ignore the following info:
A goes with T
C goes with G
You polymerize DNA 5′ –> 3′
So start at the right end of the template you typed out, starting with the primer (flip it around so that it goes 5′ –> 3′)
Then, just fill in all the way up to the C on the 5′ end of template (which will be a G on the 3′ end of coding strand).
NOW FOR THE NEW INFO:
Since you have terminators, they will halt polymerization at different location,s depending on the terminator used. For example, in the first one, you have G terminator so you can stop after adding the first G on the coding strand, or the second G, or after the third G, etc. Then just apply that same formula to each different terminator.
Hopefully, this is clear…good luck.
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