Fero-Lab-Protocols

July 25, 2018

ES and STO cell culture

By UNM CCC robin

(M. Fero — 1/4/95, with thanks to P. Soriano) MATERIALS Glassware: All hand washed with no soap. Gelatin (Sigma G-1393) Diluted to 0.1% in H2O, + 1 drop of media to colorize Plastic pipets, P100 tissue culture dishes and 24 well multiwell dishes. Trypsin (Gibco 25200-023) BioRad cuvettes (0.4 cm, electrode gap 50, Cat# 165.2088) […]

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Tags: Fero-Cell


August 2, 2016

Mouse Toe Numbering Scheme

By UNM CCC robin

Download the diagram.

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Tags: Fero-Mice


August 1, 2016

Simple Dilutions

By UNM CCC robin

Determine the final volume (Vf): How much to you want to make? Determine the dilution factor (d.f.). User either… d.f. = Cs / Cf i.e. the ratio of starting concentration / final concentration: d.f. = the fold concentration (e.g. 20x) of the starting material (if the final concentration is 1x) Calculate the starting volume (Vs): […]

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Tags: Fero-Formulae


August 1, 2016

Power Curves

By UNM CCC robin

A common issue in designing a clinical or animal study is to determine the number of subjects required. Power calculations provide an estimate of the number of study subjects that will be necessary to be able detect a difference between experimental and control subjects. The numbers required will depend on the magnitude of the difference […]

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Tags: Fero-Formulae


August 1, 2016

Formulae

By UNM CCC robin

Download the formulae here: Formulae Document

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Tags: Fero-Formulae


August 1, 2016

DirectPCR Tail DNA Preps

By UNM CCC robin

For your convenience, you can download the manual here: DirectPCR Tail DNA Preps

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Tags: Fero-Manuals


August 1, 2016

ImageJ User Guide

By UNM CCC robin

For your convenience, you can download the manual here: ImageJ-guide

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Tags: Fero-Manuals


August 1, 2016

Sorvall HB6 Rotor Specs

By UNM CCC robin

For your convenience, you can download the manual here: HB6_rotor_specs

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Tags: Fero-Manuals


August 1, 2016

Qiagen RNEasy Mini

By UNM CCC robin

For your convenience, you can download the manual here: RNeasy_Mini_Handbook

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Tags: Fero-Manuals


August 1, 2016

BioRad Bradford Protein Assay

By UNM CCC robin

For your convenience, you can download the manual here: BioRad_Bradford

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Tags: Fero-Manuals


August 1, 2016

TRIzol Reagent

By UNM CCC robin

For your convenience, you can download the manual: TRIzol_Reagent

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Tags: Fero-Manuals


August 1, 2016

Fluorescent in Situ Hybridization

By UNM CCC robin

M. Fero 4/6/2006 (courtesy of Katie Rudd, Trask lab) Materials 20 ng BAC DNA clone grown o.n. in 5 mL LB + chloramphenicol Autogen reagents Slide warmer Organics: MeOH, EtOH, formamide Fixative (3:1 MeOH:glacial HOAc) Biotin (or digoxigenin) nick-translation Kit (Invitrogen) 20x SSC (3 M NaCl, 0.3 M NaCitrate, pH 7.0) TE, pH8 CoT DNA […]

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Tags: Fero-Histology


August 1, 2016

Anti-GFP/BrdU In Situ Immunostaining

By UNM CCC robin

<R. Gu 4/6/2005 Materials 24 well dishes Millicell 0.4 µm inserts (Millipore PICM01250) – coated with fibronectin Culture medium PBS Fixative: 4% paraformaldehyde in PBS (pH 7.6) Rabbit anti-GFP (Molecular Probes, A-11122) FITC-goat anti-rabbit IgG (Jackson ImmunoResearch, 111-096-006) Biotinylated anti-BrdU (AbCam, ab2284-125) Streptavidin-Alexa Flour 555 (Invitrogen/Molecular Probes, S21381) Goat serum Horse serum BSA 2N HCl […]

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Tags: Fero-Histology


August 1, 2016

Immunostaining with Anti-Digoxigenin Alkaline Phosphatase

By UNM CCC robin

Modified from C.H. Lin 5/2005 Materials Anti-digoxigenin-AP conjugate (Roche #11093274910, 150U/200 uL, $168) Levamisole (Sigma #T1512, 2 g, $10) (blocks endogenous phosphatase activity) BCIP (Roche #11383221001, 3 mL, $49) NBT (Roche #11383213001, 3 mL, $48) Atlternate BCIP/NBT (Roche, #11697471001, 20 Tabs, makes 10 mL/Tab, $67) Blocking Reagent (Roche #11096176001, 50 g, $104, or 10x Block […]

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Tags: Fero-Histology


August 1, 2016

Immunohistochemistry (IHC) of Thymus Frozen Sections

By UNM CCC robin

(courtesy of Farr lab, UWMC) Materials Cold Acetone (-20ºC) in Coplin jars PBS, BenchKote paper, plastic lids Anti-digoxigenin-HRP diluent: 1% BSA in PBS (store 10 mL aliquots of BSA at -20ºC) Anti-rat-digoxigenin diluent: 5% Milk powder, 10% mouse serum, 10% goat serum, 1% BSA in PBS Primary and secondary antibodies (this had a link that […]

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Tags: Fero-Histology