Marccs
Badges: 4
Rep:
?
#1
Report Thread starter 5 years ago
#1
Hi
Does anyone have a simple step my step method for Sanger Sequencing for AQA BIOL 5 ?
0
reply
Serine Soul
Badges: 18
#2
Report 5 years ago
#2
From my notes:

1) To four separate tubes add:
-single stranded DNA template (sample)
-DNA polymerase
-DNA (radioactively/fluorescently) labelled primers
-Excess free nucleotides
-1 type of modified nucleotide

2) Allow tubes to undergo PCR - many copies of DNA strands formed, of various lengths (as each modified nucleotide terminates strand synthesis at different points randomly)

3) Separate fragments (one tube per well) using gel electrophoresis

4) Visualise separation using UV/X-Rays (primers were labelled so we can do this)

5) Smallest fragment moves down to the bottom so read up nucleotide by nucleotide - forming the sequence of bases complementary to original strand
1
reply
Marccs
Badges: 4
Rep:
?
#3
Report Thread starter 5 years ago
#3
(Original post by Serine Soul)
From my notes:

1) To four separate tubes add:
-single stranded DNA template (sample)
-DNA polymerase
-DNA (radioactively/fluorescently) labelled primers
-Excess free nucleotides
-1 type of modified nucleotide

2) Allow tubes to undergo PCR - many copies of DNA strands formed, of various lengths (as each modified nucleotide terminates strand synthesis at different points randomly)

3) Separate fragments (one tube per well) using gel electrophoresis

4) Visualise separation using UV/X-Rays (primers were labelled so we can do this)

5) Smallest fragment moves down to the bottom so read up nucleotide by nucleotide - forming the sequence of bases complementary to original strand
Thank You !
0
reply
X

Quick Reply

Attached files
Write a reply...
Reply
new posts
Back
to top
Latest
My Feed

See more of what you like on
The Student Room

You can personalise what you see on TSR. Tell us a little about yourself to get started.

Personalise

Poll: What factors affect your mental health most right now? Post-lockdown edition

Anxiousness about restrictions easing (45)
5.89%
Uncertainty around my education (88)
11.52%
Uncertainty around my future career prospects (80)
10.47%
Lack of purpose or motivation (98)
12.83%
Lack of support system (eg. teachers, counsellors, delays in care) (40)
5.24%
Impact lockdown had on physical health (43)
5.63%
Social worries (incl. loneliness/making friends) (83)
10.86%
Financial worries (51)
6.68%
Concern about myself or my loves ones getting/having been ill (33)
4.32%
Exposure to negative news/social media (48)
6.28%
Difficulty accessing real life entertainment (24)
3.14%
Lack of confidence in making big life decisions (72)
9.42%
Worry about missed opportunities during the pandemic (59)
7.72%

Watched Threads

View All
Latest
My Feed