User:Karmella Haynes/Notebook/PcTF Genomics/2014/05/14

From OpenWetWare

(Difference between revisions)
Jump to: navigation, search
(05/14/14)
(05/14/14)
Line 15: Line 15:
----
----
'''RT-PCR: QC_plate, all cDNA batches'''<br>
'''RT-PCR: QC_plate, all cDNA batches'''<br>
-
* Run mCh, GAPD reactions on all cDNA batches, all cell lines: 16 total
+
* Summary: Run mCh, GAPD reactions on all cDNA batches, all cell lines: 16 total. Will be used to compare cDNA quality, PcTF expression, and generate reference values
-
* Will be used to compare cDNA quality, PcTF expression, and generate reference values
+
* Experiment file name:
* Experiment file name:
-
* mCherry wells: 16x3 = 48
+
* Target gene wells: mCherry wells: 16x3 = 48; GAPD wells: 16x3 = 48
-
* GAPD wells: 16x3 = 48
+
* cDNA dilution: 1:1000 for all reactions
* cDNA dilution: 1:1000 for all reactions
 +
* Plate layout: Plate_layouts_Carly.xlsx/QC_plate
Line 42: Line 41:
-
Primer/Probe Master mixes (2 tubes)
+
Target Gene Primer/Probe Master mixes (2 tubes)
{|
{|
| align="center" style="background:#f0f0f0;"|'''Reagent'''
| align="center" style="background:#f0f0f0;"|'''Reagent'''

Revision as of 20:52, 14 May 2014

Pc-TF Genomics Main project page
Previous entry      Next entry


05/14/14

  • RT-PCR: QC_plate, all cDNA batches



RT-PCR: QC_plate, all cDNA batches

  • Summary: Run mCh, GAPD reactions on all cDNA batches, all cell lines: 16 total. Will be used to compare cDNA quality, PcTF expression, and generate reference values
  • Experiment file name:
  • Target gene wells: mCherry wells: 16x3 = 48; GAPD wells: 16x3 = 48
  • cDNA dilution: 1:1000 for all reactions
  • Plate layout: Plate_layouts_Carly.xlsx/QC_plate


cDNA Batches:

  1. K562_E001
  2. K562_C001
  3. K562_E002
  4. K562_C002
  5. K562_E003
  6. K562_C003
  7. SKNSH_E001
  8. SKNSH_C001
  9. SKNSH_E002
  10. SKNSH_C002
  11. U2OS_E001
  12. U2OS_C001
  13. U2OS_E002
  14. U2OS_C002
  15. U2OS_E003
  16. U2OS_C003


Target Gene Primer/Probe Master mixes (2 tubes)

Reagent (Single well) Gene Target mCh Gene Target GAPD (x10)
2x LC480 Probes Master(7.5 μL)360.0360.0
20 μM Forward primer(0.3 μL)14.414.4 GAPD primers
20 μM Reverse primer(0.3 μL)14.40
10 μM UPL probe(0.3 μL)14.414.4 GAPD probe
PCR H2O(0.1 μL)4.819.2
Total vol.(8.5 μL)408.0408.0


Template Master Mixes (16 tubes)

Reagent (Single well) cDNA Template no-template control
diluted Batch# cDNA(2.0 μL)12.00
PCR H2O(4.5 μL)27.00.0
Total vol.(6.5 μL)39.00.0




Personal tools