GC含量計算（GC content calcula喝畫tor）

GC content of DNA template熱好s is a critical fact員慢or which determ相劇ines the success of cloning target 嗎師genes into desired backbones.數做 Gene templates with high GC content爸器 often result in higher chances of到紙 forming self-dimers or secondary structures and require higher annealing temper票街atures.

This tool allows 志還you to determine the 好技GC content of entire明商 gene sequences as well 笑要as specific regions within a ge爸多ne. When a DNA or RNA sequence is ente問農red, the number and the percen地是tage of each base ty雜要pe is calculated. Adjusting the window 筆費size enables you to visualize the 呢唱GC content of smaller or larger segmen公頻ts within a sequence in a graphi城訊cal readout.

GC Content Calculator Crash Course in GC Content

Nucleotide bonds

In DNA and RNA, the dis房空tribution of bases has a si服鐵gnificant impact 大線on our ability 現市to study and manipulate genes. With購男in the double helix, adenine always b草空inds to thymine (or ur工電acil in RNA) using two hyd資朋rogen bonds, while guanine熱畫 binds to cytosine using three hydrogen知間 bonds (Figure 1間間). The higher the number of hydrogen b綠妹onds, the more energy require裡是d to separate the two nucleotide懂大 bases. Therefore, GC pairs w拍很ill have higher stability tha唱件n AT pairs. The perc制相entage of GC base pai跳坐rs in genomes is highly variable, b低高ut most species ran章少ge from 30-60%, with humans a要懂veraging 41%.

Figure1

Figure 1. Binding of nucleotides acr機大oss DNA strands (A),兵他 with highlighted hydrog為來en bonds for thymine-adenine pairs (B)不到 and cytosine-guanine pairs (花低C).

Applications of 國自GC content

When considering a fragment of DNA,身件 for primer desig見很n or cloning of an entire明兵 gene, it is impo話著rtant to note G木些C content given the higher amount of制頻 energy required to b很放reak the three 火雜hydrogen bonds. Polymerase chain rea白下ction (PCR) is a widely ut笑答ilized tool where GC content商短 is of high impo一訊rtance. DNA str風輛ands must be broken 地的apart (denatured), and拿下 primers must be able to bind to the 弟站correct position on the DNA 暗器strand (annealing) for amplifi秒鐵cation. The distribution of nuc飛機leotides can influence both p我機rocesses: DNA templates wit報算h high GC conten理司t may require higher場時 denaturation t懂務emperatures, and high GC content in 坐讀primers necessitates higher annealing 話笑temperatures. The optimal GC content場友 for primers is ar從小ound 50-55%, but high ampl窗低ification can be achieved wi區睡th GC content between 40 a票輛nd 60%.

Determining the GC content of an 大麗entire sequence as we窗大ll as distribution within a sequ從化ence is a valuable tool whe業校n designing and troubleshooting e民票xperiments. In addition to perfo友拿rming GC content analysis here, a體冷 GC content calculator i著用s also built into our Codon Optimization and shRNA Design tools. Both applications can be e志輛xplored on our Tools page and can be appl聽城ied to directly to 農微 your cloning experi文謝ment in the Vector Design Studio.