<> � ��� /Contents 38 0 R>> 28 0 obj The polymerase chain reaction (PCR) is a basic molecular technique used for amplifying target sequences from a DNA template in an exponential manner. x�3R��2�35W(�*T0P�R0T(�Y@���@QC= P A�J��� �14Tp�W� Therefore, it has its uses to analyze alterations of gene expression levels in tumors, microbes, or other disease states.PCR is a very powerful and practical research tool. %%EOF
Detection of potential microbial antigens by immuno-PCR (PCR-amplified immunoassay).
endobj � ��� x�3R��2�35W(�*T0P�R0T(�Y@���@QC= P A�J��� �14Sp�W� The dye does not fluoresce when in solution.PCR primers that have additional sequences on the 5′ end that become incorporated into the final PCR product.Defined region or sequence within the original DNA sample that is amplified in a PCR reaction.Machine used to rapidly and accurately shift between several temperatures in a pre-set order (for PCR).Thermophilic bacterium found in hot springs and used as a source of thermostable DNA polymerase for PCR.Enzyme that forms a polymer or string of DNA by connecting individual nucleotides.A short single-stranded piece of DNA that is complementary to the target DNA, and has a fluorophore attached to the 5′ end and a quencher attached to the 3′ end.Other PCR Techniques that are worthwhile mentioning can be listed as allele-specific PCR, ARMS (Amplification Refractory Mutation System), Assembly PCR or Polymerase Cycling Assembly (PCA), Asymmetric PCR, Dial-out PCR, Helicase-dependent amplification, Intersequence-specific PCR (ISSR), Inverse PCR, Ligation-mediated PCR, Methylation-specific PCR (MSP), Miniprimer PCR, Multiplex Ligation-dependent Probe Amplification (MLPA), Nested PCR, Overlap-extension PCR or Splicing by overlap extension (SOEing), Solid Phase PCR, Thermal asymmetric interlaced PCR (TAIL-PCR), Touchdown PCR (Step-down PCR), and Universal Fast Walking, In Silico PCR (digital PCR reactions can also be performed in the presence of labeled dNTPs or labeled primers. 131 0 obj <>
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<> The probe has a fluorophore on the 5′ end and a quencher on the 3′ end that prevents the fluorescent light from being emitted until its degraded by DNA polymerase.A fluorescent molecule that only emits fluorescent light when it is embedded or intercalated among the base pairs of double-stranded DNA.Method for using PCR to amplify unknown sequences by circularizing the template molecule.PCR reaction used specifically to amplify longer target sequences than standard PCR.
<> x�3R��2�35W(�*T0P�R0T(�Y@���@QC= P A�J��� �14Pp�W� <> endobj endobj <> Polymerase chain reaction (PCR) is the most commonly used technique in molecular biology and diagnostics. This can be caused by too low of an annealing temperature or poorly designed PCR primers that are complementary to repetitive DNA within the genome.A fluorescent probe that contains both a fluorophore and a quenching group connected by a hairpin DNA structure. 148 0 obj<>stream
<> 39 0 obj Thus, the entire PCR process can further be divided into three stages based on reaction progress: ����)h3���x7F,�Ʃ5ƿ/��@?�`��Oe$܋��\�`c���26���.��N��|������w?̗�ٺ܌Q�W�i�-���e+�؉e���(��(b�2��F�o�w�ź_��A\щY�5�W�sٍNk�I���#y�y��TR~B�R��B#��o�3r�����7�����I韮���I�$f�S5lZgv�o�������I;E)�A�ո7i�h,�{[6g�1�n���p����:c�oH��M�n5��Ϋs��zu�ގ�1��Ǯa��a;�;��s(��6Z���$&�v9E�Y� ���á�#-0aӣ��1ICWt�R� ;�n7���xy+֮Uj�bh+�廤Tj�O��J�îx�`Rvk����FY݈Gy��
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y�U��ʋ�W�`�֣���O+,�1�YG��o��S��G����\� 4̌�B�oVnmTL�h˄���_G�ǥZ�F�V�$V�nX�����?\6�l��;�����b0}w;��V�N�'�)O�����c[,��!XV6D� A���$��s�v�o��$a�{��y{$5� ���k����5��Y�n�:�� z�o2"c��d�l�#Y3��ﭵ� �V8#�+���}9�z��I H��W]��6}�� �I^�I�,�(��L�f endobj The technique is highly sensitive with the potential to produce millions to billions of copies of a specific product for sequencing, cloning, and analysis. 25 0 obj
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