User:Paulina Alatriste/Notebook/UNAM Genomics Mexico 2011/2011/06/15

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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.


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