Shared genomics buffers

The following buffers are used widely across genomics protocols and have practically infinite shelf life (all salts). All of these should be 0.22µm filter-sterilized (use syringe filters) and stored near the genomics hood in 66-219.

Note

We typically mark filtered buffers with a red Sharpie line and the word “filtered”.

  • 50 mL 1M Tris (pH 7.5)

    Component

    Concentration

    g / L

    Amount / 50 mL

    Tris-HCl

    1 M

    157.64

    7.88 g

    Elga water

    to 40 mL

    NaOH

    to pH 7.5

    ~700 uL of 12 N NaOH

    Elga water

    to 50 mL

    Filter sterilize

  • 50 mL 2M Tris (pH 7.5)

    Component

    Concentration

    g / L

    Amount / 50 mL

    Tris-HCl

    2 M

    157.64

    15.76 g

    Elga water

    to 40 mL

    NaOH

    to pH 7.5

    Elga water

    to 50 mL

    Filter sterilize

  • 50 mL 1M Tris (pH 8.0)

    Component

    Concentration

    g / L

    Amount / 50 mL

    Tris-HCl

    1 M

    157.64

    7.88 g

    Elga water

    to 40 mL

    NaOH

    to pH 8.0

    ~900 uL of 12 N NaOH

    Elga water

    to 50 mL

    Filter sterilize

  • 50 mL of 1M HEPES pH 7.5 stock solution

    Component

    Concentration

    g / L

    Amount / 50 mL

    HEPES

    1 M

    238.3

    11.915 g

    Elga water

    to 40 mL

    NaOH

    to pH 7.5

    ~1.2 mL of 12 N NaOH

    Elga water

    to 50 mL

    Filter sterilize

  • 50 mL of 500 mM EDTA (pH 8.0) stock solution

    Component

    Concentration

    g / L

    Amount / 50 mL

    EDTA

    500 mM

    186.1

    9.305 g

    Elga water

    to ~30 mL

    NaOH

    to pH 8

    ~3.75 mL of 12N NaOH

    Elga water

    to 50 mL

    Filter sterilize

    The EDTA makes the solution very acidic. Adding the NaOH will help it dissolve. It will still take a lot of vortexing to get the final solution!

  • 50 mL of 10x TE (pH 8.0):

    Component

    Concentration

    Amount / 50 mL

    2M Tris

    100 mM

    2.5 mL

    500 mM EDTA

    10 mM

    1.0 mL

    Elga water

    to ~40 mL

    36.5 mL

    NaOH

    to pH 8

    Elga water

    to 50 mL

    Filter sterilize

  • 50 mL of 5M NaCl stock solution:

    Component

    Concentration

    g / L

    Amount / 50 mL

    NaCl

    5 M

    292.21

    14.611 g

    Elga water

    to 50 mL

    Filter sterilize

  • 50 mL of 2.5M CaCl2:

    Component

    Concentration

    g / L

    Amount / 50 mL

    CaCl2 dihydrate

    2.5 M

    147.02

    18.38 g

    Elga water

    to 50 mL

    Filter sterilize

  • 50 mL of 1.0M MgCl2:

    Component

    Concentration

    g / L

    Amount / 50 mL

    MgCl2

    1.0M

    95.21

    4.76 g

    Elga water

    to 10 mL

    Filter sterilize

  • 50 mL of 10% SDS:

    Component

    Concentration

    Amount / 50 mL

    SDS

    10% (w/v)

    5 g

    Elga water

    to 50 mL