Preparation of TE-10:1 and Solution I for plasmid extraction
ABSTRACT
- Beginning of the preparation of two solutions (TE-10:1 and Solution I) needed for plasmid isolation.
- My goal of today was the preparation of two solutions for plasmid isolation, because our reserves had been finished. For that I follow the protocols that Miguel gave us. For the preparation of TE-10:1 I need the following proportions of reagents:
| Tris 1M
|
25 mL
|
| EDTA 0.25M
|
2 mL
|
| H2O HPCL
|
Take to 500 mL
|
And for the preparation of Solution I, the proportions are:
| H2O
|
100 mL
|
| Glucose 0.5M
|
11.764 mL
|
| EDTA 0.5M
|
2.353 mL
|
| Tris 1M pH 8
|
2.941 mL
|
- So the first step was to prepare the stock solutions of EDTA 0.5M pH 8, EDTA 0.25M pH 8 and glucose 0.5M. The solution of Tris 1M pH 8 was already made. So for prepare the stock I follow the next indications:
0.5M EDTA pH 8
| EDTA (disodium salt)
|
47.1 g
|
| H2O
|
Take to 250 ml
|
Adjust the pH with NaOH, filter and sterilize.
0.25M EDTA pH 8
| EDTA (disodium salt)
|
23.55 g
|
| H2O
|
Take to 250 mL
|
Glucose 0.5M
| Glucose
|
22.52 g
|
| H2O
|
Take to 250 mL
|
- I prepare this three solutions and I left in the autoclave the solutions of EDTA. Tomorrow I´m going to finish solutions TE-10:1 and Solution I.
|
UNAM Genomics Mexico 2011
