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3M Sodium Acetate (NaOAc)  

2009-07-11 06:24:39|  分类: 生物遗传天地 |  标签: |举报 |字号 订阅

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3M Sodium Acetate (NaOAc) 

From Protists and other institute

Contents

Autoclave prepared bottle for 30min on Gravity cycle

  • Be sure that we have DEPC-treated RNase-free water (in 4C walk in or -20C) or make more DEPC Water with the DEPC in the 4C drawer (be careful of built up pressure/explosion and only work with DEPC in the hood with appropriate safety protection)
  • Use barrier pipet tips and sterile (RNase-free) plasticware for this protocol

    DEPC Water (RNase-free) to weigh boat to get all crystals out and into the autoclaved bottle prepared above

  • shake like hell until all crystals are dissolved
  • add ~5.8mL glacial acetic acid (in bin under hood where acids are stored) and check on pH paper (4.7-6.8 range in live side culture materials cabinet with other pH paper) Note: this might stay at ~5.3pH for ~2min (~accurate) and then after that it changes back to dark blue, which indicates >6.8pH (this is not accurate)
  • add ~8-9 NaOH pellets to the acidic NaOAc and shake like hell until they are completely dissolved
  • Check standards of 4.0 and 7.0 buffers with the pH meter to be sure close enough, or re-standardize the machine with those two buffers (manual next to machine has instructions)
  • Put ~4mL of the well-mixed NaOAc into a 15mL orange-capped centrifuge tube (just enough to cover probe sensor)
  • Measure pH accurately using pH meter (ok if between 5.2-5.4pH, otherwise adjust with glacial acetic acid or NaOH pellets as needed)
  • Autoclave on liquid cycle for 15-20min
  • Allow to cool and aliquot into 500uL/1.5mL RNase-free tubes with labels and put in the Lab Stocks box in the -20C

    Past pH Notes for 3M NaOAc Recipe

    1. followed directions as above (up to 5.8mL glacial acetic acid from the PBJBrown glass bottle)
      1. pH = 4.8
      2. added 1 NaOH pellet and rechecked pH = 4.93
      3. added 4 NaOH pellets and rechecked pH = 5.11
      4. added 2 NaOH pellets and rechecked pH = 5.24 (ok at this)

     

    Contributor: NRH     8/97

     

    3M NaOAc pH 5.2

                  408.1g sodium acetate -3H2O

                  in 800ml H20

                  pH to 5.2 with glacial acetic acid

                  Water to 1L

                  Autoclave

    3M Sodium acetate (pH 5.2)

    AKA: NaOAc

    For 50 ml

    20.412 g

    Sodium acetate [= CH3COONa?3H2O, trihydrate]

    → ~6 ml acetic acid to pH

    → to 50 ml with dH2O

    Sodium acetate - Fisher BioReagents: BP334

    Stuart J Macdonald, 10 December 2006

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