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文献和实验Constructing a Low-budget Laser Axotomy System to Study Axon Regeneration in C. elegans
with a suitable fluorescent marker in the target neurons. Image axons at a similar magnification (0.2 um/pixel or higher magnification) and position under the crosshairs (fig.7). 7) Open the laser safety shutter and trigger the ablation laser
= 5’GCCTCCCTCGCGCCATCAGTATCGTAGGCACCTGAGASequence of 3’ primer= 5’AAAAAAAAAAAAAAAAAAAAAAAAAAAAAA/iSp18/GCCTTGCCAGCCCGCTCAGTATTGATGGTGCCTACAG(purchased from Integrated DNA Technologies, Inc)0.75 uL100 uM 5’ primer0.75 uL100 uM 3’ primer10 uL10X PCR buffer (1X = 10 mM TRIS, pH8.4, 50 mM KCl, 1.5 mM
3. Pipette the top 1 ml of supernatant to a fresh 1.5 ml microcentrifuge tube containing 0.2 ml 20% PEG/2.5 M NaCl to precipitate the phage particles. Mix by inverting several times and incubate for 15-30 minutes at room temperature. 4. Centrifuge for 15
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