Haynes Lab:Notebook/RNA-seq of PcTF Transfected U2OS & SK-N-SH cell lines/2016/08/22: Difference between revisions

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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">Today's project is...</span>
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">Today's project is...</span>
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
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==Overview==
==Overview==
Transfected K562 cells on 8/20, following [http://openwetware.org/wiki/Haynes_Lab:Notebook/RNA-seq_of_PcTF_Transfected_U2OS_%26_SK-N-SH_cell_lines/2015/06/03 same protocol as before] (from 2015-06-03).
==K562 KAH132 Results - Microscopy==
<b>24hr</b>
Well 1:<br>
[[Image:2016-08-21_Well_1_KAH132-MV2_phase_mCherry_10x.jpg | 250px]]
Well 2:<br>
[[Image:2016-08-21_Well_2_KAH132-MV2_phase_mCherry_10x.jpg | 250px]]
Well 4:<br>
[[Image:2016-08-21_Well_4_KAH132-MV2_phase_mCherry_10x.jpg | 250px]]
Well 5:<br>
[[Image:2016-08-21_Well_5_KAH132-MV2_phase_mCherry_10x.jpg | 250px]]
-----
<b>48 hr</b>
Well 2:<br>
[[Image:2016-08-22_Well_2_KAH132-MV2_phase_mCherry_10x.jpg | 250px]]
Well 5:<br>
[[Image:2016-08-22_Well_5_KAH132-MV2_phase_mCherry_10x.jpg | 250px]]
==K562 KAH132 Results - Flow Cytometry==
24 hour:<br>
[[Image:Screen_Shot_2016-09-20_at_11.10.45_AM.png]]
48 hour:<br>
[[Image:Screen_Shot_2016-09-20_at_11.10.50_AM.png]]
{| {{table}}
| align="center" style="background:#f0f0f0;"|'''Sample ID'''
| align="center" style="background:#f0f0f0;"|'''% RFP'''
|-
| 24hr A||84.8
|-
| 24hr B||84.9
|-
| 48hr A||28
|-
| 48hr B||26
|}
==K562 RNeasy prep==
Harvested cells today, RNeasy prep. Concentrations/calculations for cDNA synthesis:
{| {{table}}
| align="center" style="background:#f0f0f0;"|'''Sample ID'''
| align="center" style="background:#f0f0f0;"|'''ng/µL'''
| align="center" style="background:#f0f0f0;"|'''µL for 2 µg RNA (8 µL max)'''
| align="center" style="background:#f0f0f0;"|'''µL H2O'''
| align="center" style="background:#f0f0f0;"|'''qPCR Sample ID'''
|-
| K562 KAH132 24hr 1||917||2.2||5.8||K562_E008
|-
| K562 KAH132 24hr 2||837||2.4||5.6||K562_E009
|-
| K562 KAH132 48hr 1||879||2.3||5.7||K562_E010
|-
| K562 KAH132 48hr 2||638||3.1||4.9||K562_E011
|}
Following steps from [http://openwetware.org/wiki/Haynes:cDNA_Synthesis cDNA synthesis protocol].


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Latest revision as of 01:52, 27 September 2017

Today's project is... Main project page
Previous entry      

Overview

Transfected K562 cells on 8/20, following same protocol as before (from 2015-06-03).

K562 KAH132 Results - Microscopy

24hr

Well 1:


Well 2:


Well 4:


Well 5:


48 hr

Well 2:


Well 5:


K562 KAH132 Results - Flow Cytometry

24 hour:


48 hour:


Sample ID % RFP
24hr A 84.8
24hr B 84.9
48hr A 28
48hr B 26

K562 RNeasy prep

Harvested cells today, RNeasy prep. Concentrations/calculations for cDNA synthesis:

Sample ID ng/µL µL for 2 µg RNA (8 µL max) µL H2O qPCR Sample ID
K562 KAH132 24hr 1 917 2.2 5.8 K562_E008
K562 KAH132 24hr 2 837 2.4 5.6 K562_E009
K562 KAH132 48hr 1 879 2.3 5.7 K562_E010
K562 KAH132 48hr 2 638 3.1 4.9 K562_E011

Following steps from cDNA synthesis protocol.



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