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- Thermo scientific -fermentas
- FD0764
- 2025年09月30日
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| Catalog# | Size, concentration | Supplied with: | Certificate of Analysis | MSDS | |
| 10X FastDigest® Buffer | 10X FastDigest® Green Buffer | ||||
| FD0764 | 50 µl (50 react.) | 1.00 ml | 1.00 ml | FD0764 | |
| 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 | 20 | 2 | 80°C, 5 min | 6 |
Lambda DNA
0.7%琼脂糖
3 切割位点
1 µl FastDigest® PpuMI (Psp5II)可在
– 5分钟内酶切1 µg lambda DNA/CpoI片段;
– 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;
– 20分钟内酶切1 µg基因组DNA或120分钟内酶切5 µg基因组DNA。
Dcm: 可能重叠 – 阻止酶切。
CpG: 无重叠 – 不影响。
EcoKI: 无重叠 – 不影响。
EcoBI: 可能重叠 – 影响不确定。
| Methylation type | Sequence | Cleavage effect |
|---|---|---|
| Dcm (CCWGG) | 5'...RGGWCm5CT GG ...3' |
阻止酶切 |
| Lambda | ΦX174 | M13mp18/19 | pBR322 | puc18/19 | pUC57 |
|---|---|---|---|---|---|
| 3 | 0 | 0 | 2 | 0 | 0 |
| pTZ19R/U | pTZ57R | pBluescriptIIKS(-/+) | pBluescriptIISK(-/+) | pACYC177 | pACYC184 |
| 0 | 0 | 0 | 0 | 0 | 2 |
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文献和实验实验原理 The isolation protocol relies on base pairing between the polyA residues at the 3’ end of most mRNA, and the oligo (dT)25 residues covalently coupled to the surface of the Dynabeads® . Other RNA species lacking a polyA
E.Z.N.A.® Protocol for Bacteria
. A precipitate may form at this point. This will not interfere with RNA purification. 4. Apply sample (approximately 3.5 ml) from step 3 to an HiBind® RNA Midispin column. With the column mounted in a clean 15 ml collection tube (supplied with kit
Mag-Bind® Plasmid Mega Magnetic Protocol
. The culture density should reach 3-4 x 109 per mL. It is strongly recommended that an endA negative strain of E.coli be used for routine plasmid isolation. Examples of such strains include DH5α® and JM109® . 2. Harvest the bacterial cells
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