Daniel:Notebook/ComboLock/2017-3-21: Difference between revisions

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{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext>
{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext>
|- style="background-color:#8DB4E2;font-size:12pt;font-weight:bold" align="center"
|- style="background-color:#8DB4E2;font-size:12pt;font-weight:bold" align="center"
| width="220" height="30" | Reagent
| width="200" height="32" | Reagent
| width="85" | Single Rxn Vol
| width="120" | Single Rxn Vol
| width="90" | 8.2X MM uL
| width="120" | 14.2X MM uL


|- style="font-size:12pt"
|- style="font-size:12pt"
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|- style="background-color:#BFBFBF;font-size:12pt"
|- style="background-color:#BFBFBF;font-size:12pt"
| height="15"  valign="bottom" | Bglii Cutter Primer (10 uM)
| height="15"  valign="bottom" | Bglii Cutter Primer (10 uM)
| align="center" align="center" valign="bottom" | 0
| align="center" align="center" valign="bottom" | 5
| align="center" align="center" valign="bottom" | 0
| align="center" align="center" valign="bottom" | 71


|- style="font-size:12pt"
|- style="font-size:12pt"
| height="15"  valign="bottom" | 10X Buffer 3.1
| height="15"  valign="bottom" | 10X Buffer 3.1
| align="center" align="center" valign="bottom" | 2
| align="center" align="center" valign="bottom" | 2
| align="center" align="center" valign="bottom" | 16.4
| align="center" align="center" valign="bottom" | 28.4


|- style="background-color:#BFBFBF;font-size:12pt"
|- style="background-color:#BFBFBF;font-size:12pt"
| height="15"  valign="bottom" | Bglii
| height="15"  valign="bottom" | Bglii
| align="center" align="center" valign="bottom" | 1
| align="center" align="center" valign="bottom" | 1
| align="center" align="center" valign="bottom" | 8.2
| align="center" align="center" valign="bottom" | 14.2


|- style="font-size:12pt"
|- style="font-size:12pt"
| height="15"  valign="bottom" | nfH2O
| height="15"  valign="bottom" | nfH2O
| align="center" align="center" valign="bottom" | 12
| align="center" align="center" valign="bottom" | 7
| align="center" align="center" valign="bottom" | 98.4
| align="center" align="center" valign="bottom" | 99.4


|- style="background-color:#BFBFBF;font-size:12pt"
|- style="background-color:#BFBFBF;font-size:12pt"
| height="15"  valign="bottom" | Total
| height="15"  valign="bottom" | Total
|style="font-weight:bold" align="center" align="center" valign="bottom" | 20
|style="font-weight:bold" align="center" align="center" valign="bottom" | 20
|style="font-weight:bold" align="center" align="center" valign="bottom" | 114.8
|style="font-weight:bold" align="center" align="center" valign="bottom" | 213


|}
|}

Revision as of 16:07, 21 March 2017

Circularization Comparison (T4 Poly/Ligase; Started Wednesday)

Back to Calendar

Bgl Digestion

Because I CAN. Also, will determine if there is an improvement in signal post-digestion.

Sample Matrix

Sample Origin Condition Sample Numbers
F T4 Ligase 1,4
G Phusion/Amp (Unpurified) 1,4
H Phusion/AmpHS (Unpurified) 1,4
  1. Bglii Digestion
    1. Use samples 1 (Experiment) and 4 (No phosphate) for Bglii digest
    2. Make the following 4.2X master mix
    3. Reagent Single Rxn Vol 14.2X MM uL
      RCA Reaction Product 5 0
      Bglii Cutter Primer (10 uM) 5 71
      10X Buffer 3.1 2 28.4
      Bglii 1 14.2
      nfH2O 7 99.4
      Total 20 213
    4. Aliquot 14 uL master mix into new tubes
    5. Add 5 uL appropriate sample to each tube
    6. Use the following thermocycler program
      1. 5 min 95C
      2. 10 min 50C
      3. 1 hr 37C; When this cycle starts add 1 uL Bglii
      4. Heat kill with 65C for 20 min