OBJECTIVES:

The objective of this blog is to provide updates about what we are realizing at lab. Our work will be to repeat the procedures that we had learn in our extraordinary experience at Yale in the Mastering Metagenomics Workshop in 2010 Summer.

UPRM Team 2010

UPRM Team 2010

jueves, 28 de abril de 2011

September 27, 2010.

First of all we optimize the protocol based on the amount of sample and other materials that we had for work.

-We prepared stocks of:
CTAB  100ml and EDTA  500ml
-Computations for use 50ml bottles for the Freeze and Thaw:

Original
Modified
Total (min)
bottle
250ml
50ml

Soil
50g
15g
90g
Z buffer
75ml
22.5ml
135ml
SDS
9ml
2.7ml
16.2ml
GITC
4.5ml
1.35ml
8.1





·         The intention is to extract DNA from soil using three 50ml bottles with 15g of soil in each one.
We will have:
6 bottles=90g of soil

September 28, 2010.
-We prepared stocks of:
NaCl (5M)
Sodium Phosphate solution
Tris HCl (1M)
Z Buffer
(100ml sterile Tris, 50ml sterile Sodium Phosphate Solution, 200ml sterile EDTA, 300ml sterile NaCl, 100ml CTAB 10%, 250ml Water dH2O)
SDS 20 %
                We substitute the Lauryl with Sodium dodecyl sulfate


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