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1. BLAST: BLAST® (Basic Local Alignment Search Tool) is a set of similarity search programs designed to explore all of the available sequence databases regardless of whether the query is protein or DNA.

2. Entrez Gene: A Directory of Genes.

3. The Quantitative PCR Primer Database : (QPPD) provides information about primers and probes that can be used to quantitate human and mouse mRNA by reverse transcription polymerase chain reaction (RT–PCR) assays.

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PCR
 

The following is a basic PCR protocol. As with all experimental procedures, optimization will be required depending on the specifics of the reaction (primers, template concentration, reagents,...). For more information, see our troubleshooting guide to help solve common PCR problems.
Component
Volume
Taq DNA Polymerase 0.25 ul (0.025 u/ul)
10X Reaction Buffer 5 ul
MgCl2, 25mM
3 ul
Nuclease-Free Water complete to 50 ul
Nuclease-Free Light Mineral Oil - Do Not Autoclave 1 drop
dNTP Mix (10mM of each) 1 ul
Template DNA
 NA
Oligonucleotide Primers (upstream/downstream) NA

1. In the order listed in the table above, add the first five components to a 0.5 ml reaction tube.
2. Vortex Gently 5-10 sec.
3. Spin down the contents of the tube by brief centrifugation.
4. Add the DNA template and the primers – this will initiate the reaction.
5. Add 1 drop of mineral oil to protect against evaporation and condensation.
6. Place the reaction tubes into the thermal cycler and select the appropriate cycling reaction profile.

 

   

 

 

 

 

 

    
 

 

 

 
 
 
 
   
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