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- Thermo scientific -fermentas
- FD0944
- 2025年11月06日
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|
| Catalog# | Size, concentration | Supplied with: | Certificate of Analysis | MSDS | |
| 10X FastDigest® Buffer | 10X FastDigest® Green Buffer | ||||
| FD0944 | 20 µl (20 react.) | 1.00 ml | 1.00 ml | FD0944 | |
| Reaction temperature | Digestion time with 1µl of FastDigest® enzyme, min | bp from end of DNA required for complete digestion | Thermal inactivation | Incubation time without star activity, hours | |||
|---|---|---|---|---|---|---|---|
| Lambda, 1µg/20µl | Plasmid DNA, 1µg/20µl | PCR product, ~0.2µg/30µl | Genomic DNA, 1µg/10µl | ||||
| 37°C | 5 | 5 | 5 | 5 | 3 | 65°C, 5 min | 16 |
Lambda DNA
0.7%琼脂糖
7 切割位点
1 µl FastDigest® AclI (Psp1406I)可在
– 15分钟内酶切1 µg lambda DNA;
– 5分钟内酶切1 µg质粒DNA;
– 5分钟内酶切0.2 µg PCR扩增产物;
– 5分钟内酶切1 µg基因组DNA或30分钟内酶切5 µg基因组DNA。
Dcm: 无重叠 – 不影响。
CpG: 完全重叠 – 阻止酶切。
EcoKI: 可能重叠 – 影响不确定。
EcoBI: 可能重叠 – 影响不确定。
| Methylation type | Sequence | Cleavage effect |
|---|---|---|
| CpG | AACGTT |
阻止酶切 |
| Lambda | ΦX174 | M13mp18/19 | pBR322 | puc18/19 | pUC57 |
|---|---|---|---|---|---|
| 7 | 3 | 2 | 4 | 2 | 2 |
| pTZ19R/U | pTZ57R | pBluescriptIIKS(-/+) | pBluescriptIISK(-/+) | pACYC177 | pACYC184 |
| 2 | 2 | 2 | 2 | 2 | 2 |
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文献和实验in vitro (2).Depending on the size of the DNA to be cloned, DNA fragments are inserted into plasmid (~0.1-10kb), lambda phage (~0-23kb), or cosmid (~35-50kb) vectors. Common cloning vectors such as the pGEM® series(d), pBR322, pUC/M13, pSP series(d), pET
forpSilencer3.1-H1hygroM13F(-47)3.0REVpSilencer4_1-CMVneopSilener4.1-5'CMV-ProforpSILENCEV2.0-U6T72.0REVpSIneoT7EBVrev,T3PsitT7T7TpSK01-TM13FM13RpSK-CMV(KAN+)T7T3pSOSPsos5'Psos3'pSP64SP6M13revpSP64/65SP6此载体无反向引物pSP65SP6M13revpSP70SP6T7pSP71SP6T7pSP72SP6T7pSP73
Procedures for In Vitro Production of Bovine Embryos
after dishes are completely driedBottles/ collection medium carboy place cap loosely onto the bottle and put a piece of autoclave tape.Stir bars/ instruments place inside Tower DualPeel ® autoclave tubing and tape at both ends. Place a piece
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