University:
Swansea University
Course:
PM134 | Microbiology
Academic year:

2023
Views:

377

Pages:

2
Author:

Sanjeev K

DATE: 9.012023 18 PCR AIM To perform PCR to amplify target DNA sequence and to analyse, the ampufled DNA by agarose gel electrophoresis. PROCEDURE Add the reagents Sterile water 38 μι 10x assay buffer-5M1 10mM dNTP - Template DNA ایرا Forward primer - Iul Reverse primer - ul Taq DNA polymerase - Iul Total sreaction volume-50µl Mix all the contents. gently PCR amplification Imtial denaturation Denaturation Annealing Fina extern 94°C 94°C 48c 12°c Imin 30sec 30 cycles 30 see Extersion 12°C Imin amir Following PCR amplification add 5 ul of gel loading buffer er to each 8 Pck tubes Load 1001 of ready to use marker provided Electrophorise the sample at 100v Visualize at uv illuminator. OBSERVATION Orange coloured amplified DNA banck were Observed in lane 4, 6 and 8. DNA ladder was seen in lanel. RESULT The amplified DNA fragments at the same level confirms that target ONA has been Properly amplified.