User:Tim Henry/Notebook/Family portrait/2011/07/09

From OpenWetWare

Jump to: navigation, search
Project name Main project page
Previous entry      

Multiplex PCR attempt 1

First attempt putting multiple primers into a single reaction. Apparently this will save money and give me the results I'm looking for. The negative being that some of the primers chosen have products that are very close in size and create doublet/triplets.

Samples

  1. Tim Hair (from previous run, but accidentally left at RT)
  2. Tim Hair (new 4-5 hairs with ok roots)
  3. Gennie Hair (4-5 hairs with ok roots)

Lane Setup

Lane Setup
Lane 1 Lane 2 Lane 3 Lane 4
Sample λ-uncut TH-Old TH-New GH
Primers n/a CSF1PO, TPOX, TH01, CTLA-4 CSF1PO, TPOX, TH01, CTLA-4 CSF1PO, TPOX, TH01, CTLA-4


Aliquot new oligo: 1μL primer + 9μL dH2O into 3 tubes (for primers 7 and 8)

  • 2μL Template (forgot to add until loaded in thermal cycler)
  • 1μL each primer (8μL total)
  • 10μL dH2O

Start 1:30pm - Done 3:36pm

59 (Initial extended) didn't auto switch program to 60, so 6min at 94°C accidentally.

Results

60mL TAE + 1.2g agarose (2%)

Electrophoresis 3:49pm, 50v

End 6:40pm. Started stain, 7:08 destain

Image:07092011-2.jpg


Personal tools