Académique Documents
Professionnel Documents
Culture Documents
Aim
The aim of this experiment is to identify the absence or presence of the human
transgene in 2 samples of mouse genomic DNA using the PCR method.
1. The reagents are mixed together and centrifuged - dH 20, 10x NH4 PCR
buffer, 50 mM MgCl2, 50uM Forward primer, 50uM Reverse primer, 10mM
dNTPs, Taq polymerase (5u/ul).
2. 24ul of the mixture is aliquoted into 3 eppendorf tubes.
3. In the first tube, 1ul of genomic DNA1 was added. In the second tube 1ul
of genomic DNA2 was added and in the third tube, 1ul sterile water was
added.
4. The tubes are then left in ice to be placed into an MJ Research PCR
thermal cycler and stored at -20oC for the next part of the experiment.
5. During the next part of the experiment gel electrophoresis,
1% (w/v)
agarose gel was prepared by mixing 1g of agarose with 100mls of 1X TBE
buffer in a 250ml flask.
6. This mixture is then heated carefully in a microwave up to its boiling point
with occasional gentle swirling.
11.
5ul of Hyperladder I DNA size marker is loaded into a well and 20ul
of each of the PCR samples are loaded into the wells beside it.
12.
Results
A photographic image obtained from our group (Group 19)
Distance
migrated from
200bp
Hyperladder I
DNA molecular
weight marker
Sample 1
dH2O (Negative
Control)
(Figure 1)
(Figure 2)
Discussion
Sample 2
Distance migrated by
Sample 2 6.1cm