Covaris S2

Материал из Zbio

Перейти к: навигация, поиск

Содержание


NB! pre-PCR operation


preliminary: prepare Covaris

  • start cooling water bath[1]
  • change water in the tank
  • degas water for at least 30 min
  • check, that tank temperature is about 6-8°C


keep the pump continuously on during operation and sample processing (Degassing mode: Continious)

  1. prepare aliquots of DNA sample in LoBind tubes:
    • up to 5 µg in 120µl TE buffer, mix well
    • put aliquot into Covaris microTUBE[2] (keep the cap on the tube)
    • spin down microTUBE on strip microfuge
    • install into Covaris
  2. select run condition and start run:
    Peak [base pair]15020030040050080010001500
    Duty cycle [%]101010105552
    Intensity55443334
    Cycles per burst200200200200200200200200
    Time (seconds)4301801205580504015

  3. spin down microTUBE on strip microfuge;
  4. open cap and transfer aliquot to a new LoBind tube;
  5. optional, for several aliquots of the same sample: process next aliquot in the same microTUBE;
  6. optional, when there is enough DNA: check size distribution on the gel for each aliquot (0.05-0.15µg);
  7. optional, for several aliquots of the same sample: pool aliquots [3];
  8. snap freeze in liquid nitrogen, store at -20°C



[править] Cleaning

  • change bidestilled water daily (when in use).
  • monthly:
  1. turn off degasser;
  2. empty and wipe clean acrylic tank;
  3. wipe clean all accessible parts;
  4. fill tank with 1% bleach (1/10 dilution of 10% stock solution Na hypochlorite);
  5. turn on degasing for 10min;
  6. repeat three times:
    ○○○   empty, rinse and fill acrylic tank with normal water
    ○○○   degas for 30 sec
  7. turn on degasing for 10min;
  8. repeat three times:
    ○○○   empty, rinse and fill acrylic tank with bidistilled water
    ○○○   degas for 5 min



[править] Notes

[править] Consumables

» SnapCap microTUBE with AFA fiber and presplit Teflon/silicone/Teflon septa: Covaris, #520045 (25 tubes, ~150€)
» Holder for micro tubes: Covaris, #500114


  1. Cooling water bath:
    • pre-PCR room: VWR recirculator (set -25°C), it takes ~3h to cool down from room temperature to ~6°C (preferentially start overnight)
    • post-PCR room: Julabo waterbath, it takes ~1h to cool down from room temperature to ~6°C; (put a plastic bag with ice to accelerate cooling)
    » switch on water bath
    » switch on control unit
    » press "T"
    » press "Enter"
    » select "2.0" pressing "up/down" buttons
    » press "enter" to save
    » press "Esc"
  2. glass tubes are for better heat dissipation
  3. to be sure, that in the future all aliquots of the sample are the same
Источник — «http://molbiol.ru/wiki/Covaris_S2»


Личные инструменты


Инструменты




molbiol.ru  ·  redactor@molbiol.ru  ·  реклама

 ·  Викимарт - все интернет-магазины в одном месте  ·  Доска объявлений Board.com.ua  · 
--- сервер арендован в компании Hetzner Online, Германия ---
--- администрирование сервера: Intervipnet ---


molbiol.ru - методы, информация и программы для молекулярных биологов     Rambler's Top100 Rambler