Solution Recipes

Edit this page

Recipes for solutions and reagents needed for lab protocols.

Label your solutions

  • Please make sure solutions are labeled with your name/initials, the date that you made them

  • Please do not use stock solutions for molecular work - pour out solutions into aliquots in falcon tubes or glass jars

  • Please make sure your aliquots are labeled with the stock solution, the date they were made, who made them, and your name on the aliquot

  • Do not share aliquots with other people in the lab

7.5M ammonium acetate

Ingredient Amount
ammonium acetate (Fisher #A637500) 57.81g
water (to a final volume of 100ml)

  • If needed, sterilize by filtration in 0.2um filter.

  • Final pH will be 5.5

Bleach 10%

Ingredient Amount
Bleach 10ml
DI Water 90ml

  • DI water is located to the right of the sink in the blue lidded container with the blue valve.

  • Solution is good for up to 7 days

0.5M EDTA (pH - 8)

Ingredient Amount
Disodium ethylenediamine tetraacetate (Fisher #BP120500) 186.1g
Water 80ml
NaOH (Fisher #BP359-212) 18g

  • Stir vigorously on a magnetic stirrer.

  • Adjust the pH to 8.0.

  • Sterilize by autoclaving. (Be sure not to screw on the lid or bottle will explode)

1mM EDTA (pH - 8) for storing extracted RNA

EDTA should help stabilize and protect the RNA, but be sure to check with downstream library prep first. Use sterile 25mL pipettes to measure RNA free water and put into a 100mL glassware (autoclaved, wiped clean with RNAaseZap or RNAase Away, and rinsed with RNAase-free water). Clean magnetic spinbar with RNAaseZap. Calibrate pH meter and wipe outside with RNAase zap. Use pasteur pipette to add HCl for pH adjustment, but wipe outside with RNAase-Zap.

Ingredient Amount
Disodium ethylenediamine tetraacetate (Fisher #BP120500) 0.372 g
Water 80ml
NaOH (Fisher #BP359-212) 0.036 g
  • Stir vigorously on a magnetic stirrer. Adjust the pH to 8.0. Sterilize by autoclaving

Ethidium Bromide (10mg/ml)

Note that we are transitioning to a EtBr-free facility. CAUTION: Ethidium bromide is a mutagen and is toxic. Wear gloves when working with ethidium bromide solutions and a mask when weighting the powder.

Ingredient Amount
Ethidium bromide (Fisher #BP1025) 1g
Water 100ml

  • Stir on magnetic stirrer for several hours to ensure that the dye has dissolved

  • Light sensitive, wrap the container in aluminum foil or transfer to a dark bottle

  • Store at 4C

CTAB

Ingredient Amount
4M NaCl (Fisher #S271-1) 35 mL
0.5M EDTA pH 8.0 (Fisher #BP120500) 4 mL
1M Tris-HCl pH 8.0 10 mL
CTAB (Cetyl Trimethyl-Ammonium Bromide) (Fisher #AC22716100) 2g
  • Mix ingredients together in a clean beaker. Stir on hot plate with stir bar. Heat gently until CTAB is dissolved. Pour into graduated cylinder and bring volume up to 100ml using nanopure H2O. Pour into a bottle with a cap. Autoclave before using. Store at room temperature. Discard after 1 year.

dNTPs (10mM)

To make 48 aliquots of 25ul each of 10mM dNTPs

All of the items below come in a Fisher package #FERR01811

Ingredient Amount for 12X Amount for 1X
dTTP (100mM) 120 ul 10 ul
dATP (100mM) 120 ul 10 ul
dGTP (100mM) 120 ul 10 ul
dCTP (100mM) 120 ul 10 ul
ddH20 720 ul 60 ul

5M NaCl 500ml

Ingredient Amount
NaCl (Fisher #S271-1) 146.1 g
H2O 350 mL

*Dissolve, then bring up to volume with H2O *Sterilize by autoclaving (15 minutes)

NaOH 10N

Ingredient Amount
NaOH (Fisher #BP359-212) 40g
H20 80ml

*Continue adding water to 100ml

10% Tween-20

Ingredient Amount
100% Tween-20 (Fisher #97062-332) 1ml
Distilled H20 9 ml

TBE 10X (stock solution)

Ingredient Amount
Tris base (Fisher #BP15210) 108g
Boric acid (Fisher #A73 1) 55g *dissolve in Milli-Q water
0.5M EDTA (pH 8.0) 40ml

  • Increase the final volume to 1 liter

  • Store at room temperature or at 4C

  • Might be too concentrated for long term storage, ok if you are planning to do a lot of work

TBE 5X (stock solution)

Ingredient Amount
Tris base (Fisher #BP15210) 54 g
Boric acid (Fisher #A73 1) 27.5 g *dissolve in Milli-Q water
0.5M EDTA (pH 8.0) 20ml

  • Increase the final volume to 1 liter

  • Store at room temperature or at 4C

  • (1X TBE = 200ml of 5X TBE + 800ml)

TE

A low TE buffer that is 0.1mM EDTA is better for DNA that is destined for next-generation sequencing, because the EDTA interferes with enzymatic reactions.

Ingredient Amount
Tris-HCl (pH 8.0) X M ?
EDTA (pH 8.0) X mM ?

  • Start with a small volume of nanopure H20, about half of your final volume, in a beaker with a stirbar.

  • Measure out the amount of Tris-Hcl needed for a 10mM solution

  • Add to H20

  • Measure out the amount of EDTA needed for a 1mM solution

  • Add to H20 and Tris

  • While stirring solution, carefully insert calibrated pH probe

  • Adjust pH as needed with either HCl or NaOH

  • Note: EDTA will not fully dissolve until pH is near 8.0 and will cause pH to fluctuate as it dissolves, so be prepared to make many small adjustments in either direction ( it is best to work with diluted acids and bases).

  • Also: if you overshoot 8.0, it is okay to readjust in the other direction (even if you wildly overshoot the mark) and you DO NOT need to throw the solution out and start again

  • After pH is adjusted, transfer solution to graduated cylinder and add H20 to bring to final concentration.

  • Some people think TE buffer should not be autoclaved, but I usually autoclave it. Tris breaks down easily after autoclaving and will precipitate out more easily, so you may have to make the solution up more frequently. In my opinion, that is better than chancing a solution with DNAse.

  • Store at room temperature and discard if sediment is forming on the bottom of the container or if it starts growing or if it gets cloudy.

  • For 100ml: 1M Tris-HCl (pH 8.0) 1ml and 0.5M EDTA (pH 8.0) 200ul

  • Bring to a final volume of 100ml with nanopure H20

1M Tris

Ingredient Amount
Tris-base (Fisher #BP15210) 121.1g
  • Bring volume up to 1 liter with ddH20

1M Tris-HCL

Ingredient Amount
Tris base (Fisher #BP15210) 60.56g
ddH20 400ml
  • Adjust pH to 8.0 using HCL and bring final volume to 500ml