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Cloning Using PCR, Restriction Enzymes & ligation
Background
Related Procedures
Useful Databases & Software
Commercial Sources

Recombinant DNA lab (an interactive tutorial)

PCR Strategy (from The Protein Expression and Purification Facility of EMBL)

Let your fingers do the walking (an interactive tutorial)

Gel electrophoresis of DNA and RNA, Multiple DNA forms in gel,

DNA modifying enzymes

Setting Up a Restriction Endonuclease Reaction (NEB)

Restriction Endonucleases FAQ (NEB)

Preparing DNA
Miniprep tips (Hebrew)

PCR Cloning
1. Bio-X-act,
2. Platinum pfx,
3. Phusion

PCR Screening

1. Colony PCR screening
2.GoTaq Green Master Mix

Ligation
DNA ligation tips (Hebrew)
Direct ligation with oligo

Modifying enzymes
:
1. Klenow,
2. Mung Bean

Mutagenesis
:
1. Phusion
2. Quickchange
3. Quickchange multi

Additional Protocol Sites 
1.Molecular biology protocols from Protocol Online)
2. DNA & protein protocols
3. Molecular station protocols
4. Cold Spring Harbor Protocols

DNA Manipulation
1. Generating restrictions maps
2. Mapper
3. REbase
4. Silent mutation restriction enzyme scan 
5. Translate (nucleotides to protein) A. Expasy B. EMBOSS
6. Reverse translation (protein to nucleotides)
7. Sequence Format Conversion

PCR Design

1. Primer Tools for PCR
2. Primer Design
3. Nucleic Acids Calculators (Agilent)
4. Reverse complement
5. Oligo Calc:
6.Spectrophotometric Measurement of Nucleic Acids' Concentration 
7. Weight to Molar Quantity (for nucleic acids)  

Genetic Tools

1. REPK - finds sets of 4 commercially available restriction endonucleases which together uniquely differentiate designated sequence groups from a supplied FASTA format sequence file for use in T-RFLP.
2. PrimerZ -
primer design for genes and human SNPs (PubMed)

Tool Sets for DNA Manipulation
1. everyVECTOR
2. pDRAW32 DNA analysis software 
3. DNA Strider (for Mac)
4. EnzymeX 
5. APE - A plasmid editor
6. Sequence Manipulation Suite
7. CLC free workbench

BioSupplyNet (searching for scientific supplies.)

NEB  (restriction endo-nucleases, table of double digest,  Cleavage close to the end of DNA fragments Table 1 & Table 2 , Star activity)

Fermentas   (FastDigest restriction enzymes: Conventional restriction enzymes.)

Promega (restriction enzymes resourceCloning enzymePCR review, PCR products

Roche Applied Science (product selection )

Takara (molecular biology, PCR )

Sigma PCR product guide

Oligos Synthesis (Sigma/Genosis, Eisenberg Bros, MBC, Rhenium)



This site is maintained by Dr. Nurit Doron . Your comments are most welcome.

entries since October 2003

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