User:Allison K. Alix/Notebook/Thesis Research/2013/04/02: Difference between revisions
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Reactions of DNA with ThT and MB</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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Latest revision as of 22:35, 26 September 2017
Reactions of DNA with ThT and MB | Main project page Previous entry Next entry |
Objectives
ProceduresPart 1: Reacting DNA with MB first, followed by a reaction of the DNA-MB hybrid with AuNP 1) Prepare 727.5nM 8-repeat DNA-thiol and 800nM MB a) 727.5nM 8-repeat DNA-thiol (Original concentration of DNA is 53.1μM) 53.1μM(x)=(500μL)(1455nm) x= 13.7μL in 486.3μL H2O 1455nM(x) = 727.5nM(200μL) x= 100μL 1455nM in 100μL H2O b) Prepare 800nm MB (Original concentration of MB is 7.31μM) 7130nm(x) = (800nM)(50μL) x = 5.47μL 7.31μM in 44.53 μL H2O 3) Mix 50μL 727.5nM DNA with 50μL 800nM MB 4) Hybridize by placing mixture on heat block @ 76°C for ~25minutes 5) Turn off heat block and allow cooling to room temperature Part 2: Reacting DNA with ThT first, followed by a reaction of the DNA-ThT hybrid with AuNP 1) Prepare 727.5nM DNA (see Part 1, step 1a) 2) Prepare 800nM ThT
5mM(x) = (5μM)(10mL) 10μL of 5mM ThT in 9990μL buffer 5μM (x) = (800nm)(1mL) x= 160μL 5μM ThT in 840μL buffer 3) Mix 50μL 727.5nM DNA with 50μL 800nM ThT Part 3: React AuNP's with ethanethiol 1) Calculate how many sulfur atoms to cover 1 nanoparticle
(200x10-12m)2 = 4x10-20m2 6.1575x10-16m2/4x10-20m2 = 15393 Sulfur atoms for every 1 AuNP 2) Prepare 1mL ethanethiol solution
desired concentration of ethanethiol = 19.4nM x(15393) = 298624.2nM = 298μM density of ethanethiol = 0.839g/mL 298x10-6mol/L
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