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2. Sanger sequencing question. a). Why the Sanger sequencing chemistry would not work if too much ddNTP is added in the reaction mixtures? if we have too much ddNTP present in our reaction, we would have early termination near their 5' end and we would never generate products that are long enough to determine the sequence in the capillary electrophoresis. b) In a modified Sanger sequencing experiment, products from each reaction were recovered after separation by gel electrophoresis, and their exact molecular masses were determined by mass spectroscopy for the molecular species where all phosphodiesters are ionized. The molecular ions were found as follows: A reaction: 1163, 3050 and 3363; C reaction: 1451 and 3651; G reaction: 1779, 2107, and 2435; T reaction: 2738 and 3954. The reactions used d(AGC) as a primer. What is the sequence of the target DNA? show/explain your work.

2. Sanger sequencing question. a). Why the Sanger sequencing chemistry would not work if too much ddNTP is added in the reaction mixtures? if we have too much ddNTP present in our reaction, we would have early termination near their 5' end and we would never generate products that are long enough to determine the sequence in the capillary electrophoresis. b) In a modified Sanger sequencing experiment, products from each reaction were recovered after separation by gel electrophoresis, and their exact molecular masses were determined by mass spectroscopy for the molecular species where all phosphodiesters are ionized. The molecular ions were found as follows: A reaction: 1163, 3050 and 3363; C reaction: 1451 and 3651; G reaction: 1779, 2107, and 2435; T reaction: 2738 and 3954. The reactions used d(AGC) as a primer. What is the sequence of the target DNA? show/explain your work.

Biochemistry
Biochemistry
9th Edition
ISBN:9781319114671
Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Chapter1: Biochemistry: An Evolving Science
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2. Sanger sequencing question. a). Why the Sanger sequencing chemistry would not work if too much ddNTP is added in the reaction mixtures? if we have too much ddNTP present in our reaction, we would have early termination near their 5' end and we would never generate products that are long enough to determine the sequence in the capillary electrophoresis. b) In a modified Sanger sequencing experiment, products from each reaction were recovered after separation by gel electrophoresis, and their exact molecular masses were determined by mass spectroscopy for the molecular species where all phosphodiesters are ionized. The molecular ions were found as follows: A reaction: 1163, 3050 and 3363; C reaction: 1451 and 3651; G reaction: 1779, 2107, and 2435; T reaction: 2738 and 3954. The reactions used d(AGC) as a primer. What is the sequence of the target DNA? show/explain your work.
Transcribed Image Text:2. Sanger sequencing question. a). Why the Sanger sequencing chemistry would not work if too much ddNTP is added in the reaction mixtures? if we have too much ddNTP present in our reaction, we would have early termination near their 5' end and we would never generate products that are long enough to determine the sequence in the capillary electrophoresis. b) In a modified Sanger sequencing experiment, products from each reaction were recovered after separation by gel electrophoresis, and their exact molecular masses were determined by mass spectroscopy for the molecular species where all phosphodiesters are ionized. The molecular ions were found as follows: A reaction: 1163, 3050 and 3363; C reaction: 1451 and 3651; G reaction: 1779, 2107, and 2435; T reaction: 2738 and 3954. The reactions used d(AGC) as a primer. What is the sequence of the target DNA? show/explain your work.
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