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Goal: Compare DNA vs. RNA transfection for SARS-RBD utilizing melanoma cells and lipofectamine.
Lipofectamine Messenger MAX (LMRNA001) did not arrive in time to transfect RNA. We will continue with DNA transfection.
GFP Primers (created using cas9_eGFP)
(For Original project: CFTR - Project changed so GFP primers were not utilized)
pcDNA3-SARS-CoV-2-S-RBD-sfGFP Sequence
Linearized DNA (circular labeled: SARS-RBD-GFP-1(6/15/21 maxiprep)- 1187.43 ng/uL) with both XmaI and XhoI on 6/22, Excised on 6/23-24.
InVitro Transcription (6/24) with mMESSAGE mMACHINE T7 Ultra Kit (Freezer B)
RNA stored in -80 degree freezer (2nd floor) - CFTR Box - Row 1, Slots 5&6 - (Labeled- SARS-RBD-pcDNA3-XmaI and SARS-RBD-pcDNA3-XhoI)
DNA Gel (6/28):
Lane 4: SARS-RBD-pcDNA3-XmaI-linearization
Lane 5: SARS-RBD-pcDNA3-XhoI-linearization
(Lanes 2/3 KCNJ9 PCR: Reese - Not related to this project)
RNA Gel (6/28):
Lane 2: pcDNA3-SARS-CoV-2-S-RBD-sfGFP-XmaI
Lane 3: pcDNA3-SARS-CoV-2-S-RBD-sfGFP-XhoI - 1.5 kbp
NanoDrop RNA (6/28):
Sample 1: XmaI; Sample 2: XhoI
Trypan Blue Pre-plating (5uL trypan blue and 5uL cells):
Transfection (6/28-29):
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