连接计算器

此工具确定了在适用于典型连接反应的范围内的各种插入片段:载体摩尔比率所需的插入片段质量。

icon_0330_cc_gen_calculator-s 计算靶标输入您感兴趣的靶标的数据并接收测序计算结果
见解

 

所需的 DNA 插入片段质量 [g] = 载体 DNA 质量 [g] x {DNA 插入片段长度 / 载体 DNA 长度} x 期望的插入片段与载体摩尔比率

提示:期望的比率为集合 {1, 2, 3, 5 ,7} 中的一个

插入片段长度应小于载体长度

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首先,使用限制性内切酶消化插入片段和载体 DNA,以产生具有黏端或平端的 DNA 片段。接下来,提取期望的 DNA 片段,随后连接,然后用 DNA 连接酶进行处理。最后,将得到的完整载体转化到感受态 E. coli 细胞中,并通过合适的遗传选择方法鉴定转化体。 

 

想了解更多关于 DNA 连接的分步过程及其典型反应吗?了解更多

了解可用于连接 DNA 的酶产品

 

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了解为各种分子生物学应用设计的高品质产品。

推荐产品

T4 DNA Ligase (Rapid)
CAT NO./ID L6030-HC-L
T4 DNA Ligase (Rapid)
240,000 U of T4 DNA Ligase (600,000 U/mL) 2x Rapid Ligation Buffer and 10x T4 DNA Ligase Buffer
T7 DNA Ligase (900,000 U)
CAT NO./ID L6020L
T7 DNA Ligase (900,000 U)
900,000 U of T7 DNA Ligase (3,000,000 U/mL) and 2x Rapid Ligation Buffer
T4 DNA Polymerase (exo) (2000 U)
CAT NO./ID P7080L
T4 DNA Polymerase (exo) (2000 U)
2000 U of T4 DNA Polymerase (3000 U/mL) and 10x Blue Buffer. Storage temperature: –25°C to –15°C
T4 DNA Ligase MBG (2500 U)
CAT NO./ID EN11-250
T4 DNA Ligase MBG (2500 U)
5 x 100 μL of T4 DNA Ligase 5U/ μL, 5 x 200 μL 10x T4 Ligation Buffer, 5 x 80 μL of ATP Solution (25mM), 5 x 200 μL of 50% PEG Solution. Storage temperature: Keep at -20°C.
Klenow Fragment (2500 U)
CAT NO./ID P7060L
Klenow Fragment (2500 U)
2500 U of Klenow Fragment (0.5 mL at 5000 U/mL) and 10x Blue Buffer (1 x 1.5 mL)
Saltonase (5000 U)
CAT NO./ID EN32-050
Saltonase (5000 U)
20 µL x 250 U/µL. Storage temperature: -20°C without a defrost cycle; -80°C for long storage. Stable at -20°C for at least 24 months. No loss in activity was observed after 8 weeks of incubation at 37°C.
End-Repair Mix (low concentration)
CAT NO./ID Y9140-LC-L
End-Repair Mix (low concentration)
For 200 low-concentration DNA reactions. End-Repair Mix 1X (0.20 mL), 10X End-Repair Buffer (1 x 1.5 mL) and 1 mM dNTP solution (0.5 mL).
AllTaq PCR Core Kit (250 U)
CAT NO./ID 203123
AllTaq PCR Core Kit (250 U)
50 µl AllTaq Polymerase (5 U/µl), 1.2 ml AllTaq PCR Buffer (5x), 55 µl dNTP Mix (10 mM each), 200 µl Template Tracer (25x), 50 µl Master Mix Tracer (125x), 2 ml Q-Solution (5x), 1.2 ml MgCl2 (25 mM), 1.9 ml RNase-Free Water
Taq DNA Polymerase (250 U)
CAT NO./ID 201203
Taq DNA Polymerase (250 U)
250 units Taq DNA Polymerase, 10x PCR Buffer, 10x CoralLoad PCR Buffer, 5x Q-Solution, 25 mM MgCl2
RNase A (17,500 U)
CAT NO./ID 19101
RNase A (17,500 U)
2.5 ml (100 mg/ml; 7000 units/ml, solution) Unit definition: That amount of enzyme causing the hydrolysis of RNA at a rate such that k (velocity constant) equals unity (Kunitz units) at 25°C and pH 5.0.
HotStarTaq DNA Polymerase (250 U)
CAT NO./ID 203203
HotStarTaq DNA Polymerase (250 U)
250 units HotStarTaq DNA Polymerase, 10x PCR Buffer, 5x Q-Solution, 25 mM MgCl2
Omniscript RT Kit (50)
CAT NO./ID 205111
Omniscript RT Kit (50)
For 50 reverse-transcription reactions: 200 units Omniscript Reverse Transcriptase, 150 µl 10x Buffer RT, 100 µl dNTP Mix (contains 5 mM each dNTP), 1.1 ml RNase-free water
RNase-Free DNase Set (50)
CAT NO./ID 79254
RNase-Free DNase Set (50)
1500 Kunitz units RNase-free DNase I, RNase-free Buffer RDD, and RNase-free water for 50 RNA minipreps
QIAGEN Protease (7.5 AU)
CAT NO./ID 19155
QIAGEN Protease (7.5 AU)
7.5 Anson units per vial (lyophilized)
QIAGEN Proteinase K (2 ml)
CAT NO./ID 19131
QIAGEN Proteinase K (2 ml)
2 ml (>600 mAU/ml, solution)