User:Karmella Haynes/Notebook/BioBrick cloning/2015/01/22: Difference between revisions
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">Karmella's BioBrick Cloning</span> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;">Karmella's BioBrick Cloning</span> | ||
|style="background-color: #F2F2F2" align="center"| | |style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} | ||
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== | ==01/22/15== | ||
<!-- Precede finished items with a checkmark ✓ --> | <!-- Precede finished items with a checkmark ✓ --> | ||
* | * Gal4 fusion cloning | ||
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Try different cDNA libraries for the ATF2 fragment to account for any mutations or deletions | Try different cDNA libraries for the ATF2 fragment to account for any mutations or deletions | ||
# KAH60 "backbone" (without VP64, includes scars) = 6637: KAH60 F1/ KAH60 R1 | # KAH60 "backbone" (without VP64, includes scars) = 6637: KAH60 F1/ KAH60 R1 | ||
# U2OS ATF2 insert = 906: ATF2_f1/ ATF2_r1 | # U2OS C002, 1:1000 cDNA dilution; ATF2 insert = 906: ATF2_f1/ ATF2_r1 | ||
# SKNSH | # SKNSH C001, 1:1000 cDNA dilution; same | ||
# K562 | # K562 C001, 1:1000 cDNA dilution; same | ||
{| {{table}} cellspacing="3" <!-- | {| {{table}} cellspacing="3" <!-- PCR rxn. table --> | ||
|- valign="top" | |- valign="top" | ||
| bgcolor=#cfcfcf | Reagent | | bgcolor=#cfcfcf | Reagent | ||
| bgcolor=#cfcfcf | Rxn1 | | bgcolor=#cfcfcf | Rxn1 | ||
| bgcolor=#cfcfcf | Rxn2 | | bgcolor=#cfcfcf | Rxn2 | ||
| rowspan="7" | < | | bgcolor=#cfcfcf | Rxn3 | ||
| bgcolor=#cfcfcf | Rxn4 | |||
| rowspan="7" | Expected:<br>1. KAH60 = 6637<br>2. ATF2 = 906<br>3. ATF2 = 906<br>4. ATF2 = 906 | |||
| rowspan="7" | [[Image:KAH012315_gel.jpg|270px|Hover name]]<br>10 μL/lane, 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] | |||
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| Template || | | Template || 0.2 || 1.0 || 1.0 || 1.0 | ||
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| 10 uM fwd primer || 1.0 || 1.0 || 1.0 || 1.0 | | 10 uM fwd primer || 1.0 || 1.0 || 1.0 || 1.0 | ||
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| 2x GoTaq green || 12.5 || 12.5 || 12.5 || 12.5 | | 2x GoTaq green || 12.5 || 12.5 || 12.5 || 12.5 | ||
|- | |- | ||
| dH<sub>2</sub>O || | | dH<sub>2</sub>O || 10.3 || 9.5 || 9.5 || 9.5 | ||
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| || 25 | | || 25.0 || 25.0 || 25.0 || 25.0 | ||
|} | |} | ||
Program: GOTAQ35cyc | |||
* 95°C, 3 min | |||
* 30x[95°C, 1 min; 57°C, 1 min; 72°C, 3 min] | |||
* 72°C, 3 min | |||
* 4°C ∞ | |||
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Latest revision as of 00:40, 27 September 2017
Karmella's BioBrick Cloning | Main project page Previous entry Next entry | |||||||||||||||||||||||||||||||||||||
01/22/15
Planning: Gal4 fusion cloning
Program: GOTAQ35cyc
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