Protocols

DNA Preparation - from Bacteria

DNA-Miniprep. - Rapid boiling
My favorite protocol to prep. DNA from E.coli, if you have more than 12 samples its faster than a commercial kit.

DNA-Miniprep - Alkaline Lysis
My favorite protocol to prep. DNA from Agrobacterium, it is also great for E. coli.

DNA-Miniprep - Alkaline Lysis for BACs
My favorite protocol to prep. BACs. It is has some slight modifications to the previous protocol.

DNA-Miniprep - Quick & Dirty Alkaline Lysis
A very fast alkaline lysis protocol. It is popular because you just pour over the solvents.

DNA-Maxiprep - Alkaline Lysis
My favorite protocol for maxipreps. It takes its time, but you get huge amounts of DNA.

DNA Preparation

DNA Miniprep using CTAB
The best protocol to get big amounts of DNA from plant material.

RNA Preparation

Trizol combined with Columns
The best protocol to get a lot of RNA which is also clean.

RNA Miniprep using hot phenol
A dangerous protocol to extract RNA from all kinds of tissues.

RNA Miniprep using CTAB
The best protocol to get big amounts of RNA from plant material.

RNA Miniprep using 2-Butoxyethanol
The best protocol to get RNA from material rich in phenoles, sugars or oxidizing compounds.

RNA Midiprep using 2-Butoxyethanol
The best protocol to get RNA from material rich in phenoles, sugars or oxidizing compounds.

General Cloning Methods

Ligation
How to make a successfull ligation. This is a basic protocol.

Separate RNA and DNA
My favourite protocol to get pure DNA or RNA - or both! It just workes.

cDNA synthesis using superscript II
My favourite protocol for cDNA synthesis. Takes a little but workes perfectly.

Creating an artificial DNA fragment (Dimer)
How to make a DNA fragment with a sequence of your choise. Usefull to create multiple cloning sites.

Cloning with Gateway Technology (based on Invitrogen's System)

TOPO-Reaction
How to make a successfull TOPO reaction with pENTR-D/Topo vector from invitrogen.

Gateway BP-Reaction II
How to make a pENTR clone from a PCR product and a pDONR clone.

Classic LR-Reaction II
How to make your final clone from one pENTR clone and one pDEST clone.

Multiple Gateway Reaction II without Mastermix
How to make your final clone from three pENTR clones and one pDEST clone.

Multiple Gateway Reaction II using a Mastermix
How to make your final clone from three pENTR clones and one pDEST clone.

Competent Cells

Starter Glycerol Stocks
How to make starter glycerol stocks to speed up competent cell production.

Preparation of Chemical Competent Cells
How to make good chemical competent cells.

Transformation of Chemical Competent Cells
How to transform chemical competent cells.

Preparation of Electro-Competent Cells
How to make good electro-competent cells.

Transformation of Electro-Competent Cells
How to transform electro-competent cells.

Adneoviral Vectors

Cloning of Adenoviral Vectors
Some things to consider if you start cloning an adenoviral vector.

Production of Adenoviral Vectors
How to produce adenoviral vectors starting from a plasmid.

Titration of Adenoviral Vectors
How to titrate adenoviral vectors to figure out their concentration.

Production of Adenoviral Vectors for Application in Test Animals
A good method to produce adenoviral vectors for test animals.

Cloning with the first generation Gateway Technology

Old gateway protocols using the first version of the enzymes, were buffer and enzyme were provided separate. Due to the lower stability I would not recomend to use this protocols.

Gateway BP-Reaction
How to make a pENTR clone from a PCR product and a pDONR clone.

Classic LR-Reaction
How to make your final clone from one pENTR clone and one pDEST clone.

Multiple Gateway
How to make your final clone from three pENTR clones and one pDEST clone.

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