This lab was performed in partners. To begin with the experiment, the gel caster was assembled. The two plates were properly cleaned and then introduced well align to avoid leakages to the plastic platform. Once there were correctly adjusted. A wash bottle was used to pour water to the edges and wait for about 5 minutes to verify there was no leakage. The TA came to the group in order to verify the correct assembly. Then, the water was through on the sink.
40 ml of 15% resolving gel solution was prepared. A 100 ml beaker was selected and labeled as resolving gel solution. To the beaker 9.68 ml of water was added plus 20 ml of 30% Acrylamide. Once all the solution were added to the beaker it was let aside in order to prepared the 4x resolving …show more content…
20 ml of 5% stacking gel solution was prepared. A 100 ml beaker was selected and labeled as stacking gel solution. To the beaker 11.5 ml of water was added plus 3.3 ml of 30% Acrylamide. Once all the solution were added to the beaker it was let aside in order to prepared the 4x stacking gel buffer needed to be added to the beaker. To make the desired buffer, another 100 ml beaker was used to prepare a solution buffer of 50 ml. Following the prelab calculations once again and using the weighting balance, the amount of Tris and SDS needed was weighted. The Tris used were 3.206g and SDS 0.205g. The solids were added to the beaker and to this 50 ml of water. The beaker was stir on the stirring plate until the solution dissolved completely. The already calibrated pH meter was used to measure he initial pH of the buffer as 10.40. HCl was added drop by drop in order to lower the pH to the desired 6.8. A final pH of 6.62 was achieved. Once the buffer was ready, 5 ml of it were added with the graduated cylinder to the initial beaker with the stacking gel. The TA then added the rest to the solution, these being 20 microliters of TEMED and 300 microliters of 10% APS. These last two materials were added right before pouring the gel to the gel
40 ml of 15% resolving gel solution was prepared. A 100 ml beaker was selected and labeled as resolving gel solution. To the beaker 9.68 ml of water was added plus 20 ml of 30% Acrylamide. Once all the solution were added to the beaker it was let aside in order to prepared the 4x resolving …show more content…
20 ml of 5% stacking gel solution was prepared. A 100 ml beaker was selected and labeled as stacking gel solution. To the beaker 11.5 ml of water was added plus 3.3 ml of 30% Acrylamide. Once all the solution were added to the beaker it was let aside in order to prepared the 4x stacking gel buffer needed to be added to the beaker. To make the desired buffer, another 100 ml beaker was used to prepare a solution buffer of 50 ml. Following the prelab calculations once again and using the weighting balance, the amount of Tris and SDS needed was weighted. The Tris used were 3.206g and SDS 0.205g. The solids were added to the beaker and to this 50 ml of water. The beaker was stir on the stirring plate until the solution dissolved completely. The already calibrated pH meter was used to measure he initial pH of the buffer as 10.40. HCl was added drop by drop in order to lower the pH to the desired 6.8. A final pH of 6.62 was achieved. Once the buffer was ready, 5 ml of it were added with the graduated cylinder to the initial beaker with the stacking gel. The TA then added the rest to the solution, these being 20 microliters of TEMED and 300 microliters of 10% APS. These last two materials were added right before pouring the gel to the gel