连接计算器

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.

€179.00

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)

€450.00

CAT NO./ID EN32-050

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 12 months. No loss in activity was observed after 8 weeks of incubation at 37°C.

€134.00

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).

€474.00

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

€209.00

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

€164.00

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.

€323.00

CAT NO./ID 203203

HotStarTaq DNA Polymerase (250 U)

250 units HotStarTaq DNA Polymerase, 10x PCR Buffer, 5x Q-Solution, 25 mM MgCl2

€216.00

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

€364.00

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

€143.00

CAT NO./ID 19155

QIAGEN Protease (7.5 AU)

7.5 Anson units per vial (lyophilized)

€108.00

CAT NO./ID 19131

QIAGEN Proteinase K (2 ml)

2 ml (>600 mAU/ml, solution)

€114.00

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。

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

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。