DNA spiking, also known as custom spiking, is the differing ratio of bases at a single degenerate position when synthesizing oligonucleotides. DNA spiking is a unequal proportions of bases at a given position (for example, 10% Adenine, 75% Guanine, 5% Cytosine & 10% Thymine). As an example, with the degenerate code R = A + G, 50% of the time that R position is adenine and the other 50% of the time it is guanine. However, with DNA Spiking, the R position could be adenine 70% of the time and guanine 30% of the time. The proportions do not need to be 70:30, the ratios can be anything else such as 12:82 and 64:36.[1]
DNA spiking can also refer to a spike control in PCR, which is when DNA is added to a sample that will provide some signal (e.g. a plasmid or some synthetic DNA with a specific known sequence) to a reaction, and seeing if the reaction will amplify. This method is used to discover if the PCR method is working correctly, as in a PCR machine it may not amplify DNA properly, so by adding spiked DNA it can be observed how much DNA is produced. This is then compared to the amount of DNA that would be theoretically predicted if the machine was working properly so that any malfunctions can be discovered.[2]
^"Can somebody explain to me what "spiking" means in RT-PCR and why do you - General Lab Techniques". www.protocol-online.org. Retrieved 2017-10-07.
January 01, 1970
spiking, also, known, custom, spiking, differing, ratio, bases, single, degenerate, position, when, synthesizing, oligonucleotides, unequal, proportions, bases, given, position, example, adenine, guanine, cytosine, thymine, example, with, degenerate, code, tim. DNA spiking also known as custom spiking is the differing ratio of bases at a single degenerate position when synthesizing oligonucleotides DNA spiking is a unequal proportions of bases at a given position for example 10 Adenine 75 Guanine 5 Cytosine amp 10 Thymine As an example with the degenerate code R A G 50 of the time that R position is adenine and the other 50 of the time it is guanine However with DNA Spiking the R position could be adenine 70 of the time and guanine 30 of the time The proportions do not need to be 70 30 the ratios can be anything else such as 12 82 and 64 36 1 DNA spiking can also refer to a spike control in PCR which is when DNA is added to a sample that will provide some signal e g a plasmid or some synthetic DNA with a specific known sequence to a reaction and seeing if the reaction will amplify This method is used to discover if the PCR method is working correctly as in a PCR machine it may not amplify DNA properly so by adding spiked DNA it can be observed how much DNA is produced This is then compared to the amount of DNA that would be theoretically predicted if the machine was working properly so that any malfunctions can be discovered 2 See also editRNA spike inReferences edit Degenerate Bases amp Spiking Introduction www genelink com Retrieved 2017 10 07 Can somebody explain to me what spiking means in RT PCR and why do you General Lab Techniques www protocol online org Retrieved 2017 10 07 Retrieved from https en wikipedia org w index php title DNA spiking amp oldid 1070551051, wikipedia, wiki, book, books, library,
