Total RNA Purification Kit - Tissue

Random Hexamers are short oligodeoxyribonucleotides of random sequence [d(N)6]. Primers are quality controlled (MS), purified (reversed phase HPLC), lyophilised and aliquoted and came with a guaranteed amount of at least 5OD (about 76.1nmol (136.4µg)). Hexanucleotide primers are a mixture of random 5'-hydroxyl hexanucleotides or hexamers, and can be used to quickly and efficiently prepare radioactive or non-radioactive probes using a DNA polymerase and a suitable DNA template or for cDNA synthesis from mRNA. The heterogeneous nature of the random primers ensures that all possible sequences will be represented in the probe mixture. As a result, these primers are well suited for generating as long as possible (complete) transcripts of the RNA, transcribing the 5 'ends, in particular for long RNA templates. As an alternative you can also use Oligo(dT)-Primer > for reverse transcription. Oligo(dT) primer bind to the polyA tail of RNA and therefore transcription will start always at the 3'-end of the RNA. The guaranteed delivery quantity of 5OD, corresponding to approx. 76.1nmol, or 136.4μg, can be dissolved in 760μL DEPC water to give a concentration of 100pmol/μL (100μM), or approx. 180μg/μL. If using between 50ng and 250ng of the hexamers per 20μL reverse transcription reaction, 760μL (100μM) will be sufficient for 460 to 2280 reverse transcriptase reactions.

The Genaxxon M-MuLV Reverse Transcriptase, encoded by Moloney Murine Leukemia Virus (M-MuLV RT) and expressed in E.coli is an RNA-dependent DNA polymerase lacking the RNase H activity. The enzyme synthesizes the cDNA first strand from a single-stranded RNA template to which a primer has been hybridized. M-MuLV RT will also extend primers hybridized to single-stranded DNA! The enzyme is recommended for synthesis of cDNA from 100 bp up to 10 kb length. Second strand cDNA synthesis can be achieved from some mRNA templates without an additional DNA polymerase. Genaxxon offers the HotScriptase master mixes (M3062 >; M3064 >) with which it is possible to synthesize double stranded DNA completely without an isothermal transcription step. Time saving: up to 50%! No need for Superscript!Money saving: high! With our high quality dNTPs as Set (M3015.4100 and M3015.0250) or Mix (M3016.1010) or our DNA Ladders and our favourable standard agarose (M3044) we can offer additional high quality products for your PCR.

Ceramic beads for the lysis of tissue from a wide range of sample material, e.g. for RNA extraction. The kit contains 100 preps with 1.4mm ceramic beads (zirconia) and comes pre-proportioned to 1g in 2mL standard screw cap tubes. The beads are suitable for the homogenization of "soft" tissue from lung, liver, brain, kidney, spleen, skin, cells from cell culture, as well as for plant tissues such as e.g. Leaves or even whole insects. The usabl sample amount ranges from up to 200mg tissue, up to 200μL of cells suspended in water or isotonic saline solution, or up to 100mg wet weight of tissue culture cells grown in suspension (e.g., ≤10E7 mammalian cells). The beads can be used for a variety of samples and applications, e.g. B. for veterinary diagnostics (BSE screening), pharmacology and toxicology (drug detection), criminology and forensic medicine, food industry (GMO detection), microbiology in the environment (microorganism lysis). as well as the isolation of nucleic acids and proteins. They are chemically inert and do not bind nucleic acids. The beads are comparable with MP Biomedical No. 116913100 (equivalent to Lysing Matrix D) and are compatible with most commercially available homogenizers, like FastPrep® (MP Biomedicals), MagNA Lyser Instrument (Roche). You can reorder our RNA > and DNA purification columns > for the isolation of total RNA or DNA from enzymatic reactions or ceramic beads > separately. These articles can also be used with buffers/kits from other suppliers. With our HotScriptase RT PCR master mix >, you can quickly and easily transcribe and amplify RNA into DNA by normal PCR. You do not need a reverse transcriptase and no reverse transcription step. 

Oligo (dT)20 are single stranded oligodeoxyribonucleotides containing only deoxythymine (dT) to be able to prime with the poly(A) tail of mRNA molecules. Primers are quality controlled (MS), purified (reversed phase HPLC), aliquoted and lyophilized and came with a guaranteed amount of at least 5OD (about 27.2nmol (163.7µg)). Oligo (dT)20 Primer is designed to initiate the synthesis of cDNA from total RNA in a reverse transcription reaction, where Reverse Transcriptase, e.g. MMuLV (M3042) > is starting the reaction from the poly-A-end of mRNA. It can also be used for generation of labeled cDNA to screen microarrays. A mixture (1:1 ratio) of random hexamer primers (M3038 >) and Oligo(dT)20 primer may improve the sensitivity of cDNA synthesis as it will especially improve the efficiency of transcripts longer than 600 bp. Guaranteed delivery of at least 5OD (equals about 27.2nmol (163.7µg)). Primers are quality controlled (MS), purified (reversed phase HPLC), lyophilised and aliquoted. Dissolve content (5 OD) in 271.5μL of molecular biology grade water for an end concentration of 100 pmol/μL (100μM). 760μL (100μM) are sufficient for 760 reverse transcriptions of 20μL each. Oligo (dT)20 Primer is suitable for use as a primer for first strand cDNA synthesis with a reverse transcriptase, such as MMuLV or AMV. The primer hybridizes to the poly-adenylated tail found on the 3´ end of most eukaryotic mRNAs. Oligo (dT)20 ensures that the 3´ end of mRNAs are represented. Oligo(dT)20 Primer may be better suited for the new generation of Reverse Transcriptases like SuperScript™ that work at higher temperatures as the longer Oligo(dT)20 Primer enable annealing in reverse transcription reactions at higher temperatures. Applications• cDNA synthesis from total RNA in a reverse transcription reaction.• optimal choice for construction of cDNA libraries from eukaryotic mRNAs.• Full length cDNA cloning• 3’ rapid amplification of cDNA ends (3’ RACE)• Generation of labelled cDNA for screening of microarrays.Oligo (dT)20 Primer can not be used together with degraded RNA, prokaryotic RNA or miRNA (lack of poly(A) tail). Primer Sequence: 5´ – d (TTT TTT TTT TTT TTT TTT TT) –3´

Lysozyme (Muramidase) preferentially hydrolyses the β-1,4-glycosidic binding between N-Acetyl muraminic acid and N-Acetyl glucosamine, a component of the proteoglycan-cell wall of certain microorganisms. The enzyme is present in many organisms. In molecular biology, the enzyme from chicken white egg is used to lyse E. coli for the isolation of plasmid-DNA. The working concentration is 200-300 μg/µL. To increase the plasmid yield (approx. 5-10%) lysozyme may be added. Another application is the lysis of bacteria for the preparation of bacterial RNA. In this case the working concentration is 40μg/mL (stock solution 50mg/mL). Stability: The lyophilized powder of lysozyme is stable for many years at +2°C to +8°C. In solution, the stability at pH values from 4 to 5 at +2°C to +8°C is several weeks and at room temperature several days. pH-optimum is at 9.2, the isoelectric point at 11.0. Lysozyme will be inhibited by surfactants like SDS or alcohols and fatty acids, imidazole and indol-derivatives.