Oligonucleotide Probe Design and Synthesis

Design oligonucleotide probes by looking for an area of the sequence that matches the order for the selected allele but for no other group.[8] A combination of sense and antisense sequences can be used to formulate the micro-array for low-resolution HLA-DQA1 typing (Fig. 1). When developing and choosing optimal probes, take into consideration: 1) desired probe length: the optimal probe length is 12 bases, which provides a high level of discrimination between alleles and allows hybridization to take place at room temperature; 2) placement of the mismatched base(s): optimal placement of the mismatched bases is near the center of the sequence; 3) suitable G/C content: G/C content should range from 42% to 58%; and 4) free energy values: values of free energy should be greater than — 3.1 AG so that the duplex formations are kept to a minimum. Manual design is not very successful. The best results are obtained by using

DQA1*010101 ACT GCC TGG CGG TGG CCT GAG TTC AGC AAA TTT GGA GGT TTT GAC CCG CAG

DQA1*010102 ---------------------------------------------------

DQA1*010201 --- --- --- --- --- --- --- --- --- --- --- --- --- --- --- --- ---

DQA1*010202 --- --- --- --- --- --- --- --- --- --- --- --- --- --- --- --- ---

DQA1*0103 --- --- --- --- --- --- --- --- --- --- --- --- --- --- --- --- ---

DQA1*010401 ---------------------------------------------------

DQA1*010402 ---------------------------------------------------

DQA1*0105 ---------------------------------------------------

DQA1*0106 G-- --- --- --- --- --- --- --- --- --- --- --- --- --- --- --- ---

DQAl*02a DQAl*02b

5 ' -GTC TGG AAG TTG- 3 ' 5 ' -C CAC AGA CTT AG- 3 '

DQA1*030101

DQA1*0302

DQA1*0303

Getting Started With Dumbbells

Getting Started With Dumbbells

The use of dumbbells gives you a much more comprehensive strengthening effect because the workout engages your stabilizer muscles, in addition to the muscle you may be pin-pointing. Without all of the belts and artificial stabilizers of a machine, you also engage your core muscles, which are your body's natural stabilizers.

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