Difference between revisions of "User:Karmella Haynes/Notebook/PcTF Genomics/2012/12/18"

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(Autocreate 2012/12/18 Entry for User:Karmella_Haynes/Notebook/PcTF_Genomics)
 
(mm/dd/yy)
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==mm/dd/yy==
 
==mm/dd/yy==
 
<!-- Precede finished items with a checkmark &#x2713; -->
 
<!-- Precede finished items with a checkmark &#x2713; -->
* Line item 1
+
* TriZOL-RNeasy prep: lyse cells in TriZOL; store at -80°C
  
 
----
 
----
'''Line item 1'''<br>
+
'''TriZOL-RNeasy prep'''<br>
> Samples
+
# "K562 1": 1:2 dilution cells, +DNA
 
+
# "K562 1": 1:2 dilution cells, no DNA
{| class="wikitable" border="0" cellspacing="3" <!-- Rxn. table -->
+
# "K562 1": 1:4 dilution cells, +DNA
|-valign="top"
+
# "K562 1": 1:4 dilution cells, no DNA
| <u>Reagent</u> || <u>Volume</u>
+
# "K562 1": 1:8 dilution cells, +DNA
|-
+
# "K562 1": 1:8 dilution cells, no DNA
| reagent 1 || # μL
 
|-
 
| reagent 2 || #
 
|-
 
| reagent 3 || #
 
|-
 
| reagent 4 || #
 
|-
 
| dH<sub>2</sub>O || #
 
|-
 
| &nbsp; || # μL
 
|}
 
 
 
--> Reaction conditions
 
  
 +
* Collect 1 mL cells from 10 cm plate into 2.0 mL microfuge tubes (4x); discard remainder of culture
 +
* Pellet cells at r.t./ 3 min./ 2000 rpm
 +
* Discard supernatant
 +
* Resuspend pellet in 500 μL TriZOL
 +
* Store at -80°C for next steps (Carly)
  
  

Revision as of 15:05, 18 December 2012

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mm/dd/yy

  • TriZOL-RNeasy prep: lyse cells in TriZOL; store at -80°C

TriZOL-RNeasy prep

  1. "K562 1": 1:2 dilution cells, +DNA
  2. "K562 1": 1:2 dilution cells, no DNA
  3. "K562 1": 1:4 dilution cells, +DNA
  4. "K562 1": 1:4 dilution cells, no DNA
  5. "K562 1": 1:8 dilution cells, +DNA
  6. "K562 1": 1:8 dilution cells, no DNA
  • Collect 1 mL cells from 10 cm plate into 2.0 mL microfuge tubes (4x); discard remainder of culture
  • Pellet cells at r.t./ 3 min./ 2000 rpm
  • Discard supernatant
  • Resuspend pellet in 500 μL TriZOL
  • Store at -80°C for next steps (Carly)