Difference between revisions of "Sequence DNA for Bio113"

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(Sequencing Your Plasmid DNA to Confirm Cloned DNA Sequence)
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'''Sequencing Primers to confirm Inserts v2.0'''<br>
 
'''Sequencing Primers to confirm Inserts v2.0'''<br>
 
113_pClone_SeqFor<br>
 
113_pClone_SeqFor<br>
CTAATTCAACAAGAATTGGGAC Tm = 60 C  65bp upstream first base<br>
+
CTAATTCAACAAGAATTGGGAC Tm = 60° C  65bp upstream first base<br>
 
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<br>
 
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<br>
 
113_rClone_SeqFor <br>
 
113_rClone_SeqFor <br>
AACGTGCTGAAGGTCGTC Tm = 65 C  60bp upstream first base<br>
+
AACGTGCTGAAGGTCGTC Tm = 65° C  60bp upstream first base<br>
 
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<br>
 
<br>
 
113_actClone_SeqFor<br>
 
113_actClone_SeqFor<br>
GCATTAGAAACCGTCCATCG Tm = 64 C 73bp upstream first base<br>
+
GCATTAGAAACCGTCCATCG Tm = 64° C 73bp upstream first base<br>
 
113_actClone SeqRev<br>
 
113_actClone SeqRev<br>
CCATTAGAAACCATCCCTCG Tm = 63 C 107bp upstream first base<br>
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CCATTAGAAACCATCCCTCG Tm = 63° C 107bp upstream first base<br>
 
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<br>
 
<br>
 
<br>
 
113_repClone_SeqFor<br>
 
113_repClone_SeqFor<br>
ACAGCTCTTCGCCTTTAC  Tm = 62 C 93bp upstream first base<br>
+
ACAGCTCTTCGCCTTTAC  Tm = 62° C 93bp upstream first base<br>
 
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Revision as of 21:24, 6 November 2017

Sequencing Your Plasmid DNA to Confirm Cloned DNA Sequence

  1. For each eXperimental sample, determine the volume of DNA you need to deliver 320 ng of DNA to a barcoded sequencing tube. Record the bar code for each sample.
  2. Add water to your DNA until the combined volume is 8 µL.
  3. To each of your 8 µL of DNA, add 4 μL of the appropriate sequencing primer (at 2 µM concentration) for a total volume of 12 μL.
  4. Record your sample and group name for each barcoded tube you used. You should have 3 new eXperimental and one old eXperimental sequencing reactions to send away. Those working with actClone will have 8 tubes to send away.



Sequencing Primers to confirm Inserts v2.0
113_pClone_SeqFor
CTAATTCAACAAGAATTGGGAC Tm = 60° C 65bp upstream first base


113_rClone_SeqFor
AACGTGCTGAAGGTCGTC Tm = 65° C 60bp upstream first base


113_actClone_SeqFor
GCATTAGAAACCGTCCATCG Tm = 64° C 73bp upstream first base
113_actClone SeqRev
CCATTAGAAACCATCCCTCG Tm = 63° C 107bp upstream first base


113_repClone_SeqFor
ACAGCTCTTCGCCTTTAC Tm = 62° C 93bp upstream first base