"
Team:Bordeaux/NotebookJune4
From 2014.igem.org
(Difference between revisions)
(Created page with "{{:Team:Bordeaux/Tete}} <html> <h1>2<sup>nd</sup> June </h1> <div class="type-exp"> Molecular biology</div> <br> <ul class="day-detail"> <li> <div> <b>DAY THREE :</b> Grow cel...") |
|||
| (One intermediate revision not shown) | |||
| Line 2: | Line 2: | ||
<html> | <html> | ||
| - | <h1> | + | <h1>4<sup>th</sup> June </h1> |
<div class="type-exp"> Molecular biology</div> | <div class="type-exp"> Molecular biology</div> | ||
| Line 10: | Line 10: | ||
<li> | <li> | ||
<div> <b>DAY THREE :</b> Grow cells until A600: 0.5 – 0.7</div><br> | <div> <b>DAY THREE :</b> Grow cells until A600: 0.5 – 0.7</div><br> | ||
| - | <div> <b>Subsequent steps should be carried out in the cold room on ice : </b>Put flask on ice for 10 minutes, then spin cells down at 2500xg (3350 RPM) in JLA 8.1 rotor. Pour off supernatant. Resuspend gently in 80 mL – I first resuspend in 25 mL TB, then add remaining 295 mL. Leave on ice for 10 minutes; spin down again at 2500 xg for 10 minutes. Resuspend cells in 20 mL TB, add 1.4 mL of DMSO dropwise while gently shaking [Final DMSO concentration is 7%]. Aliquot in 100 μL aliquots (you’ll need about 450 pre-chilled 0.5 mL tubes). Flash freeze in liquid nitrogen. Store at -80°C (the lower the rack the better).</div><br> | + | <div> <b>Subsequent steps should be carried out in the cold room on ice : </b><br>Put flask on ice for 10 minutes, then spin cells down at 2500xg (3350 RPM) in JLA 8.1 rotor. Pour off supernatant. Resuspend gently in 80 mL – I first resuspend in 25 mL TB, then add remaining 295 mL. Leave on ice for 10 minutes; spin down again at 2500 xg for 10 minutes. Resuspend cells in 20 mL TB, add 1.4 mL of DMSO dropwise while gently shaking [Final DMSO concentration is 7%]. Aliquot in 100 μL aliquots (you’ll need about 450 pre-chilled 0.5 mL tubes). Flash freeze in liquid nitrogen. Store at -80°C (the lower the rack the better).</div><br> |
</li> | </li> | ||
</ul> | </ul> | ||
Latest revision as of 19:54, 17 October 2014
4th June
Molecular biology
-
DAY THREE : Grow cells until A600: 0.5 – 0.7
Subsequent steps should be carried out in the cold room on ice :
Put flask on ice for 10 minutes, then spin cells down at 2500xg (3350 RPM) in JLA 8.1 rotor. Pour off supernatant. Resuspend gently in 80 mL – I first resuspend in 25 mL TB, then add remaining 295 mL. Leave on ice for 10 minutes; spin down again at 2500 xg for 10 minutes. Resuspend cells in 20 mL TB, add 1.4 mL of DMSO dropwise while gently shaking [Final DMSO concentration is 7%]. Aliquot in 100 μL aliquots (you’ll need about 450 pre-chilled 0.5 mL tubes). Flash freeze in liquid nitrogen. Store at -80°C (the lower the rack the better).
