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    April 80 Scallop QuantSeq
    • Notes
    • Metadata
    • Relevant Items
    • RNA Extraction - Trizol and PureLink Kit
    • QuantSeq 3' mRNA Kit
    Editing disabled on read-only entry.

     
    April 80 Scallop QuantSeq

    Wednesday, 4/11/2018180416-Pfister-Silliman_Eukaryote_Total_RNA_Nano_2.pdf180416-Pfister-Silliman_Eukaryote_Total_RNA_Nano_1.pdfSaturday, 4/14/2018Sunday, 4/15/2018180416-Pfister-Silliman_Eukaryote_Total_RNA_Nano_1.pdfMonday, 4/16/2018Wednesday, 4/18/2018Thursday, 4/19/2018Scallop_QS_042018.xlsxFriday, 4/20/2018Sunday, 4/22/2018Monday, 4/23/2018Tuesday, 4/24/2018Wednesday, 4/25/2018Thursday, 5/3/2018Friday, 5/4/2018180504_Pfister-Silliman_HS_KS-Lib5-80.pdf
    Wednesday, 4/11/2018
    Extractions, 8 using Enzymax columns and 8 using RNeasy columns
    180416-Pfister-Silliman_Eukaryote_Total_RNA_Nano_2.pdf
    180416-Pfister-Silliman_Eukaryote_Total_RNA_Nano_1.pdf
    Saturday, 4/14/2018
    Extractions
    Sunday, 4/15/2018
    Extractions
    Dissect: 50_M, 50_C, 63_M, 63_C
    180416-Pfister-Silliman_Eukaryote_Total_RNA_Nano_1.pdf
    Monday, 4/16/2018
    Dissections of 23 scallop tissues, left in -80 afterwards for extraction on Wed:
    364C, 246C, 44C, 392C, 375C, 48C, 236C, 66M, 240C, 364M, 48M, 39M, 66C, 375M, 246M, 44M, 392M, 377M, 377C, 240M, 236M, 364M, 242M, 363M_2
    Wednesday, 4/18/2018
    Extractions in 2 sets (morning and afternoon). All Enzymax columns
    236M: column looked funky
    all Enzymax columns
    2nd round: 48M, 246M, 39M, 66M, 363M_2, 240M, 392M, 246C, 240C
    Thursday, 4/19/2018
    Working sheet for calculating uL to pool, etc:
    Scallop_QS_042018.xlsx
    Round 1 to Step 4.
    1 set of Bioanalyzer
    Friday, 4/20/2018
    Concentrate RNA:
    15uL: 58M, 311M, 312M, 361M
    10uL: 386M, 37M, 242G, 39G, 45G
    7uL: 363M (pooled both), 236M
    Up to qPCR step of Round 1
    Sunday, 4/22/2018
    Made qPCR mix, did not run until Monday (8:30a)
    Monday, 4/23/2018
    Ran qPCR on 32 samples from Round 1. NTC showed some flourescence...contamination? Because it was left over night? Will run full PCR based on estimated PCR cycles from qPCR and do thorough Bioanalyzer...
    Did Round 2 through 1st strand synthesis. Mightttt have flipped the tubes in each row. Labelled as is, will check.
    Tuesday, 4/24/2018
    Round 2 through 2nd strand synthesis
    Round 1: PCR, Qubit
    Wednesday, 4/25/2018
    Round 1: Bioanalyzer of 22 samples
    2SS purification, qPCR of Round 2
    Thursday, 5/3/2018
    ●
    Checked qPCR of Round 2.
    ●
    Ran PCR of Round 2.
    ●
    Re-amplified the 5 samples from Round 1 that did not work.
    ●
    PCR purification in Round 2 and RE samples.
    ●
    Quibit-> had to use AccuGreen HS kit by Biotium (annoying), so re-Qubitted 12 of the Round 1 samples to see if values are close.
    ○
    Concentrations were generally higher (might be from evaporation too? So redoing all samples with Biotium Qubit
    Friday, 5/4/2018
    Bioanalyzer of RE samples and some from Round 1.
    Library submitted!
    Bioanalyzer of pool from Genomics Core:
    180504_Pfister-Silliman_HS_KS-Lib5-80.pdf

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