AlanFung:LabNotes/Capturing/2014-1-2: Difference between revisions

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>Alan6017518
(Created page with "==Helping Dheeraj to test Klentaq capture reactions== *Dheeraj reached out to me saying he wants to start a qPCR reaction on the Klentaq capture reaction, he was working with ...")
 
>Alan6017518
Line 3: Line 3:
*They have 12 samples and we will be amplifying the captured samples with qPCR using the same protocol as it in Stoffol capture
*They have 12 samples and we will be amplifying the captured samples with qPCR using the same protocol as it in Stoffol capture
==Protocol==
==Protocol==
==PCR amplification to check if capture works==
{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8<\hiddentext>
|- style="font-size:12pt;font-weight:bold"  valign="bottom"
| width="137" height="15" | Components
| width="65" | 1x rxn (ul)
| width="65" | 12x rxn (ul)
| width="65" | Final Conc.
|- style="font-size:12pt"  valign="bottom"
|style="font-weight:bold" height="15" | Circularized DNA template
| align="center" | 2.5
| align="center" | 0
| align="center" | &nbsp;
|- style="font-size:12pt"  valign="bottom"
|style="font-weight:bold" height="15" | KAPA SSYBR FAST MM 2X
| align="center" | 12.5
| align="center" | 150
| 1X
|- style="font-size:12pt"  valign="bottom"
|style="font-weight:bold" height="15" | 10uM AmpF6.4Sol
| align="center" | 0.5
| align="center" | 6
| 200nM
|- style="font-size:12pt"  valign="bottom"
|style="font-weight:bold" height="15" | 10uM AmpR6.3IndX
| align="center" | 0.5
| align="center" | 6
| 200nM
|- style="font-size:12pt"  valign="bottom"
|style="font-weight:bold" height="15" | H2O
| align="center" | 10
| align="center" | 120
| align="center" | &nbsp;
|- style="font-size:12pt"  valign="bottom"
|style="font-weight:bold" height="15" | Total
| align="center" | 25
| align="center" | &nbsp;
| align="center" | &nbsp;
|}

Revision as of 21:21, 2 January 2014

Helping Dheeraj to test Klentaq capture reactions

  • Dheeraj reached out to me saying he wants to start a qPCR reaction on the Klentaq capture reaction, he was working with Noi to test the protocol, but Noi is sick and I will be helping to perform the qPCR
  • They have 12 samples and we will be amplifying the captured samples with qPCR using the same protocol as it in Stoffol capture

Protocol

PCR amplification to check if capture works

Components 1x rxn (ul) 12x rxn (ul) Final Conc.
Circularized DNA template 2.5 0  
KAPA SSYBR FAST MM 2X 12.5 150 1X
10uM AmpF6.4Sol 0.5 6 200nM
10uM AmpR6.3IndX 0.5 6 200nM
H2O 10 120  
Total 25