PDF4PRO ⚡AMP

Modern search engine that looking for books and documents around the web

Example: bachelor of science

Making Electrocompetent Cells

Making Electrocompetent Cells Day 1. 1. Streak out frozen glycerol stock of bacterial Cells (Top 10, DH5 , etc.) onto an LB. plate (no antibiotics). Grow plate overnight at 37 C. Day 2. 1. Autoclave: 2 L of ddH2O. 100 mL of 10% v/v glycerol (molecular biology grade). 1 L LB (or your preferred media). 4 centrifuge bottles and caps Lots of microfuge tubes 2. Chill overnight at 4 C: ddH2O. 10% glycerol Centrifuge rotor 3. Prepare starter culture of Cells Select a single colony of E. coli from fresh LB plate and inoculate a 10 mL starter culture of LB (or your preferred media). Grow culture at 37 C in shaker overnight. Notes: You can also substitute other media like SOB, 2xYT, etc for LB if preferred. All glassware should be detergent free. Trace detergent residue reduces competency. Day 3. 1. Inoculate 1 L of LB media with 10 mL starter culture and grow in 37 C shaker.

3. Add 10g Bacto Yeast Extract. 4. Add 5g NaCl. 5. Adjust pH to 7.0 with 5N NaOH. 6. Adjust to 1L with distilled H2O. 7. Sterilize by autoclaving on LIQUID cycle. LB (Luria-Bertani) Broth Recipe 10 grams tryptone 5 grams of yeast extract 10 grams of NaCl (NOTE: A premixed powder is also available from Fisher cat# BP1426) 1. Dissolve in 1 L of ...

Fullscreen Download
Loading..

Tags:

  Extracts, Cabots, Yeast, Yeast extract, Electrocompetent, Yeast extract bacto

Information

Domain:

Source:

Link to this page:

Please notify us if you found a problem with this document:

Spam in document Broken preview Other abuse

Transcription of Making Electrocompetent Cells

Documents from same domain

cyclic-AMP (cAMP) cascade protein kinase A (PKA). …

mcb.berkeley.edu

The cyclic-AMP (cAMP) cascade begins with the production of cAMP, which is a pure signaling molecule, i.e., not a metabolism intermediate. cAMP binds and activates protein kinase A (PKA). PKA converts phosphorylase-b into phosphorylase-a, which is phosphorylated and active, in two steps. PKA converts glycogen

  Mapc

PowerPoint Presentation - ALZHEIMER’S DISEASE

mcb.berkeley.edu

Alzheimer’s Disease Progresses Through Distinct Stages Mild Moderate Severe Memory loss Language problems Mood swings Personality changes Diminished

  Disease, Presentation, Alzheimer, Alzheimer s disease

NEUROBIOLOGY LABORATORY MCB 160L

mcb.berkeley.edu

Neurobiology is a very diverse field. The nervous system is studied by an incredible variety of techniques in electrophysiology, biochemistry, anatomy, cell biology, molecular biology,

  L160, Neurobiology, Mcb 160l

GLYCOLYSIS REVIEW & OVERVIEW

mcb.berkeley.edu

The “committed step”: fructose 6-phosphate fructose 1,6-bisphosphate. Two triose compounds are isomerized and oxidized to retrieve ATP & NADH via glyceraldehydes 3-phosphate dehydrogenase. The pathway concludes with a strong, irreversible, ATP …

  Phosphate, Dehydrogenase, Glycolysis, Phosphate dehydrogenase

PENTOSE PHOSPHATE PATHWAY

mcb.berkeley.edu

Metabolism Lecture 5 — PENTOSE PHOSPHATE PATHWAY — Restricted for students enrolled in MCB102, UC Berkeley, Spring 2008 ONLY Oxidative Phase: Glucose-6-P Ribose-5-P Glucose 6-phosphate dehydrogenase.

  Pathway, Phosphate, Dehydrogenase, Stepone, Phosphate dehydrogenase, Pentose phosphate pathway

A quick guide to sketching phase planes

mcb.berkeley.edu

A quick guide to sketching phase planes Section 6.1 of the text discusses equilibrium points and analysis of the phase plane. However, there is one idea, not mentioned in the book, that is very useful to sketching and analyzing phase planes, namely nullclines. Recall the basic setup for an autonomous system of two DEs: dx dt = f(x,y) dy dt = g(x,y)

  Panels

Practice problems answ

mcb.berkeley.edu

Practice problems (with answers) This is the degree of difficulty of the questions that will be on the test. This is not a practice test because I did not consider how long it would take to finish these problems. It also does not have a matching section, which I will include on the test. 1. DNA polymorphisms on the Y chromosome and on mtDNA ...

  Practices

I Introduction II G Protein-coupled Receptors III Receptor ...

mcb.berkeley.edu

Signaling 5 MCB110-Spring 2008-Nogales In paracrine signaling the signaling molecule affects only target cells in the proximity of the signaling cell. An example is the conduction of an electric signal from one nerve cell to another or to a muscle cell. In this case the signaling molecule is a neurotransmitter. In autocrine signaling cells ...

  Signaling

cyclic-AMP (cAMP) cascade protein kinase A (PKA). PKA

mcb.berkeley.edu

Metabolism Lecture 7 — METABOLIC_REGULATION— Restricted for students enrolled in MCB102, UC Berkeley, Spring 2008 ONLY Phases of the TCA Cycle. [1] Acetyl-CoA production: Organic fuels Acetyl-CoA [2] Acetyl-CoA oxidation: Acetyl-CoA enters TCA and is enzymatically oxidized, but energy is conserved in electron carriers, NADH FADH 2

  Regulations, Fuel

A Guide to Crimping RJ-45 Connectors

mcb.berkeley.edu

RJ-45 connectors intended for CAT-5 cable to attach properly to CAT-6 cable. When you're shopping for RJ-45 connectors, make sure the ones you buy are suitable for CAT-6. The picture below shows a CAT-5 and a CAT-6 connector the CAT-6 connector is the darker one).

  Connectors, Crimping