Items Tagged ‘Fero-RNA’

June 1st, 2016

384 Well Template

By UNM CC UNM CC

Download the pdf file.

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June 1st, 2016

RNA Gel Electrophoresis

By UNM CC UNM CC

Matthew Fero, 4/5/06 (see Molecular Cloning 7.21-7.34) Materials DEPC H2O DEPC 0.1% (v/v) q.s. de-ioinized H2O 37ºC x1 hr, or r.t. overnight Autoclave (NaOAc, EDTA and ethidium bromide solutions should also be DEPC treated. Tris has a reactive amine and can’t be DEPC treated) 10x Formaldehyde gel loading buffer: 50% glycerol 10 mM EDTA, pH8 […]

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June 1st, 2016

mRNA Amplification with T7 RNA Polymerase

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M. Fero 12/05 Materials MessageAmp II aRNA Amplication Kit (Cat #1751, Ambion) 100% Ethanol RNA samples (e.g. 1 µg RNA per sample) Procedure Reverse transcription Verify that EtOH (24 mL) has been added to the Wash Buffer. Turn on a 42ºC oven and set PCR machine to hold at 70ºC. In an Eppendorf tube add: […]

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June 1st, 2016

RNEasy Midi Protocol

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M. Fero — 6/01 Materials Qiagen RNEasy Midi Kit Dounce homogenizers (for tissues) Syringes and needles (for cultured cells) 70% EtOH Sterile 15 mL conical tubes Procedure Before using the reagents for the first time add 10 µL of ßME per mL of Buffer RLT, and add 4 vol of EtOH to buffer RPE. For […]

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May 18th, 2016

TRIzol Prep

By UNM CC UNM CC

v.2 M. Fero 6/1/06 Procedure Homogenize cells (10 million) or tissue (50-100 mg) in 1 mL TRIzol Reagent (e.g. scrape and pass through 30G needle, dounce homogenize and pass through needle, or use a homogenizer) and transfer to a 1.5 mL tube. Optional – Spin at 12k g for 10′ at r.t. to pellet debris. […]

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May 18th, 2016

Bioanalyzer Sample Preparation

By UNM CC UNM CC

1. Prepare a 3 µL aliquot of each RNA sample at a concentration of 50 – 500 ng/uL in RNAse free H2O. 2. Submit to Genomics Core (cost = $4 / sample) See Bioanalyzer manual for more details.

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May 18th, 2016

Amino-Allyl cDNA Labeling

By UNM CC UNM CC

M. Fero. v.4 Materials RNA sample 30 µg Total or 2 µg mRNA Oligo dT (dT18VN) 5 µg/µL, 34.8 (µg/mL)/A260 Superscript II, 5x 1st strand buffer (Invitrogen) 1 M DTT (Sigma, 43816, $16.80/10 mL) 50x dNTP Mix (dATP, dCTP, dGTP @25 mM, TTP=15mM) 10x AA-dUTP, 2 mM (MW = 523 g/mol), (Sigma A0410, $193.50/1mg – […]

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Tags: Fero-RNA