What is SuperScript IV?

Unit definition. One unit of SuperScript™ IV RT is the amount of enzyme required to incorporate 1 nmole of deoxyribonucleotide into acid-precipitable material in 10 min at 37°C using poly(A) oligo(dT)12-18 as a template/primer.

What is RT MMLV?

M-MLV Reverse Transcriptase is a recombinant DNA polymerase that synthesizes a complementary DNA strand from single-stranded RNA, DNA, or an RNA:DNA hybrid. Compared to AMV RT, Moloney Murine Leukemia Virus Reverse Transcriptase (M-MLV RT) lacks DNA endonuclease activity and has a lower RNase H activity.

How do random hexamer primers work?

Random hexamer primers bind throughout the entire length of RNA, ensuring reverse transcription of all RNA sequences due to their random structure. A mixture of both random hexamer and oligo(dT) is possible, as well. The third choice is a gene-specific primer.

What is M MuLV?

Moloney Murine Leukemia Virus (M-MuLV, MMLV) Reverse Transcriptase is an RNA-directed DNA polymerase. This enzyme can synthesize a complementary DNA strand initiating from a primer using either RNA (cDNA synthesis) or single-stranded DNA as a template.

What is an Oligo dT primer?

Description. Oligo (dT)18 Primer is single-stranded sequence of deoxythymine (dT), used for priming reactions catalysed by reverse transcriptase. The transcript is primed in the poly(A) tail of mRNA molecules.

What is the difference between Oligo dT and random primers?

Oligo(dT) primers amplify only mRNAs containing a poly(A) tail, since that is where the primer binds to promote reverse transcription. Random primers amplify most RNA species, including degraded RNA and viral genomes.

How do you make tris buffer solution?

To prepare base solvent add 3ml 20% SDS to add 3.75mL 1M Tris buffer at pH 6.8 in a suitable container. Add 9 mg bromphenol blue, 1.16 gm DTT (or 2.4ml B-mercaptoethanol) and mix well. Add 4.5mL glycerol to the solution, mix well.

How do you prepare a western blot loading buffer?

Contributed by Martin Fitzpatrick, University of Birmingham, United Kingdom 15ml stock solution of western blot loading buffer. Dilute for use. To prepare base solvent add 3ml 20% SDS to add 3.75mL 1M Tris buffer at pH 6.8 in a suitable container.

How do you make a base solvent from SDS?

To prepare base solvent add 3ml 20% SDS to add 3.75mL 1M Tris buffer at pH 6.8 in a suitable container. Add 9 mg bromphenol blue, 1.16 gm DTT (or 2.4ml B-mercaptoethanol) and mix well.

What is the best buffer to use for reverse transcription?

5x first strand synthesis buffer. For use in reverse transcription. Temperature sensitive items must be shipped overnight. Use the form below to request a Certificate of Analysis.