Difference between revisions of "20.109(S08): TA notes for module 1"
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#*order # 28104, 50 rxns | #*order # 28104, 50 rxns | ||
#*1 rxn per group | #*1 rxn per group | ||
− | #pCX-NNX, | + | #pCX-NNX, 20 μL per group |
#NEB2 Buffer (~5 μL used per group) | #NEB2 Buffer (~5 μL used per group) | ||
#EcoRI and XbaI enzymes (~2 μL used per group) | #EcoRI and XbaI enzymes (~2 μL used per group) | ||
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#*pCX-NNX, XbaI cut only | #*pCX-NNX, XbaI cut only | ||
#*pCX-NNX, EcoRI cut only | #*pCX-NNX, EcoRI cut only | ||
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#TC Room | #TC Room | ||
#*Razors, Face Masks, Film/Camera, Transilluminator, Spatulas, Kimwips, Eppendorf, Pen, EtBR waste jar | #*Razors, Face Masks, Film/Camera, Transilluminator, Spatulas, Kimwips, Eppendorf, Pen, EtBR waste jar | ||
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*Run purified products (2 per group) on an agarose gel at the end of the day - post photograph. | *Run purified products (2 per group) on an agarose gel at the end of the day - post photograph. | ||
*Freeze DNA at end of day. | *Freeze DNA at end of day. | ||
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'''Prep Work''' | '''Prep Work''' | ||
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'''Day of Lab:''' | '''Day of Lab:''' | ||
+ | |||
+ | *Students Benchtop | ||
+ | :*LB in 15mL conical tube | ||
+ | :*LB+Amp plates (5 per group, so at least 30 per lab) | ||
+ | :*1mL 3M NaAc | ||
+ | :*Ice buckets with ice | ||
+ | :*15mL conical tube with 100% EtOH in ice | ||
+ | :*Check spreaders, refill EtOH beakers and alcohol burners | ||
+ | |||
+ | *Instructors Benchtop | ||
+ | :*Ice bucket with ligation buffer | ||
+ | :*tRNA | ||
+ | :*2x 50mL conical tube with 70% EtOH | ||
+ | :*pCX-EGFP transformation control | ||
+ | :*Midway through lab, thaw supercomp cells (~400-500ul/group, need 12 tubes of ~200ul each for class of 12) | ||
+ | |||
*Quiz (prepared by TA). | *Quiz (prepared by TA). | ||
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#FACS tubes | #FACS tubes | ||
#*16 per group, plus extras | #*16 per group, plus extras | ||
− | #*BD Falcon tubes only, VWR cat # 60819-310 | + | #*BD Falcon tubes only, VWR cat # 60819-310 (1000x) or 60819-295 (500x) |
'''Day of Lab:''' | '''Day of Lab:''' | ||
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===Agarose Gel=== | ===Agarose Gel=== | ||
− | # DNA gel: 1% agarose gel in 1X TAE, 1 g agarose, 100mL 1X TAE, 2 | + | # DNA gel: 1% agarose gel in 1X TAE, 1 g agarose, 100mL 1X TAE, 2 |