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RNAspectra FDD Kits (for Fluorescent Differential Display)
Product Description
For Fluorescent Differential Display with three fluorescently labeled one-base anchored oligo-dT primers and rationally designed arbitrary 13mers.
The RNAspectra Kits feature a new generation of Differential Display technology based on fluorescent detection of PCR products instead of radioactivity. It is the first non-radioactive differential display system with similiar sensitivity to that of 33P isotopic labeling.
RNAspectra Red Kit 1 | |
1. 5X Reverse Transcription Buffer | 200 µL |
2. MMLV Reverse Transcriptase (100 u/µL) | 40 µL |
3. RH-T11G Anchor Primer | 200 µL |
4. RH-T11A Anchor Primer | 200 µL |
5. RH-T11C Anchor Primer | 200 µL |
6. dNTP Mix (FDD) | 500 µL |
7. 10X PCR Buffer | 500 µL |
8. H-AP1 Arbitrary Primer | 150 µL |
9. H-AP2 Arbitrary Primer | 150 µL |
10. H-AP3 Arbitrary Primer | 150 µL |
11. H-AP4 Arbitrary Primer | 150 µL |
12. H-AP5 Arbitrary Primer | 150 µL |
13. H-AP6 Arbitrary Primer | 150 µL |
14. H-AP7 Arbitrary Primer | 150 µL |
15. H-AP8 Arbitrary Primer | 150 µL |
16. Control RNA (1 µg/µL) | 1 µL |
17. FDD Loading Dye | 0.5 mL |
18. H-T11G Anchor Primer | 300 µL |
19. H-T11A Anchor Primer | 300 µL |
20. H-T11C Anchor Primer | 300 µL |
Each kit contains all the reagents needed for fluorescent differential display except RNA samples, Taq DNA polymerase, and a fluorescent scanner. The MessageClean® is also highly recommended prior to differential display to remove trace amounts of DNA contamination from RNA samples.
GenHunter currently sells 3 versions of the RNAspectra Kit: a "Red", a "Green" and a "Yellow" kit. As in the RNAimage® Kit, the RNAspectra kit contains three one-base-anchored oligo-dT primers to subdivide the mRNA population, except they are labeled with either 5' Rhodamine (RNAspectra Red Kits), 5' Fluorescein (RNAspectra Green Kits), or 5' NED (RNAspectra Yellow Kits). The use of three one-base-anchored primers reduces the redundancy and potential under-representation of the sub-population of mRNAs encountered by using either twelve individual or four degenerate two-base-anchored oligo-dT primers. With built-in restriction sites at the 5' ends of both anchored and arbitrary 13mers, the longer primer pairs produce highly selective and reproducible cDNA patterns. These improvements ensure that differentially expressed genes are more readily identified, cloned, and manipulated.
We also offer the RNAimage Differential Display Kits for standard isotopic DD.
All kits are shipped on dry ice via overnight delivery. A detailed step-by step protocol is included.
RNAspectra® Kits 2-10 contain all the components of Kit 1, but each has 8 different arbitrary 13mers (H-AP9 to H-AP80).
One could start with any RNAspectra® Kit, since the sequences are random and designed to be maximally different from one another, but to ensure a maximum coverage, multiple kits may be needed.
The RNAspectra® kits have been optimized for use with the Hitachi FMBIO® II or III Fluorescence Imaging System. For more information contact us at 800-311-8260 or info@genhunter.com
RNAspectra Red Kit 1 | |
1. 5X Reverse Transcription Buffer | 200 µL |
2. MMLV Reverse Transcriptase (100 u/µL) | 40 µL |
3. RH-T11G Anchor Primer | 200 µL |
4. RH-T11A Anchor Primer | 200 µL |
5. RH-T11C Anchor Primer | 200 µL |
6. dNTP Mix (FDD) | 500 µL |
7. 10X PCR Buffer | 500 µL |
8. H-AP1 Arbitrary Primer | 150 µL |
9. H-AP2 Arbitrary Primer | 150 µL |
10. H-AP3 Arbitrary Primer | 150 µL |
11. H-AP4 Arbitrary Primer | 150 µL |
12. H-AP5 Arbitrary Primer | 150 µL |
13. H-AP6 Arbitrary Primer | 150 µL |
14. H-AP7 Arbitrary Primer | 150 µL |
15. H-AP8 Arbitrary Primer | 150 µL |
16. Control RNA (1 µg/µL) | 1 µL |
17. FDD Loading Dye | 0.5 mL |
18. H-T11G Anchor Primer | 300 µL |
19. H-T11A Anchor Primer | 300 µL |
20. H-T11C Anchor Primer | 300 µL |
Each kit contains all the reagents needed for fluorescent differential display except RNA samples, Taq DNA polymerase, and a fluorescent scanner. The MessageClean® is also highly recommended prior to differential display to remove trace amounts of DNA contamination from RNA samples.
Kit Cat. Nos. | Red Kit | Green Kit | Yellow Kit |
RNAspectra Kit 1 | R501 | F501 | Y501 |
RNAspectra Kit 2 | R502 | F502 | Y502 |
RNAspectra Kit 3 | R503 | F503 | Y503 |
RNAspectra Kit 4 | R504 | F504 | Y504 |
RNAspectra Kit 5 | R505 | F505 | Y505 |
RNAspectra Kit 6 | R506 | F506 | Y506 |
RNAspectra Kit 7 | R507 | F507 | Y507 |
RNAspectra Kit 8 | R508 | F508 | Y508 |
RNAspectra Kit 9 | R509 | F509 | Y509 |
RNAspectra Kit 10 | R510 | F510 | Y510 |
We also offer the RNAimage Differential Display Kits for standard isotopic DD.