User:Tara K. Luckau/Notebook/Team ConGen/2011/01/11

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Gel Touchdown

 * ---[[Image:20110111_GelSchematicTD.jpg|580 px]]
 * [[Image:20110111_Touchdown.TIF|600 px]]
 * Faint bands at very bottom, near next rows' lanes = primer front
 * ladder triplet: 396/344/298
 * ladder doublet: 220/201
 * ladder doublet: 154/134
 * ladder band: 75
 * but no amplicon at expected 450bp     [[Image:sad-face.jpg|50 px]]

Gel Timber Rattlesnakes

 * --[[Image:20110111_GelSchematicTR2.jpg|280 px]]
 * [[Image:20110111_TR2.TIF|300 px]]


 * ---[[Image:20110111_GelSchematicTR10.jpg|460 px]]
 * [[Image:20110111_TR10.TIF|485 px]]


 * Definitely bands! Too bad I ran off primer 10's bands (but I assume they're there)

Next Steps

 * Try to break the PCR by using Tara's dNTPs, Tara's Buffer
 * Try to make Scun PCRs work by using Rulon's dNTPs, 10x Buffer + MgCl2

Extract Sceloporus undulatus

 * acquired from Tod Reeder; "H0074; TWR 78" (looks like muscle and liver tissue)
 * extract same species as primers were designed for, to rule out DNA extraction and primer synthesis as PCR problem
 * 1) Add 300µL of Cell Lysis Solution to about 10mg tissue
 * 2) Add 2µL Proteinase K (20mg/mL); incubate overnight at 55°C water bath


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