What is the efficacy of cDNA synthesis ?

23 December 2016 1 1K Report

dear all

I extracted RNA from cell line and measured my RNA by nanodrop then I used 1st strand cDNAsynthesis kit to synthesize cDNA and measured the cDNA by using nanodrop ssDNA ,I used 1000ng/ul of RNA but I get between 600 to 700 of cDNA?

my question is it usual to get 60-70% cDNA

Rafah Al-Nafakh

Thanks a lot for your answer, I didn't clean up my cDNA or RNA I just treated the RNA with DNase . my 260/280and 260/230 was between 1.8 to 2 for all the samples,I just get worried when I saw the difference between RNA and c DNA concentration because I divided my RNA samples into 3 groups and synthesized the cDNA in3 consecutive days so the 1st group I got like 90 to 95% cDNA ,while the 2nd and the 3rd groups got just 60 to 70% cDNA

Badges
Science topic

More Rafah Al-Nafakh's questions See All

Can we distinguish the type of free fluid inside the pelvic and abdominal cavities whether blood or serous fluid using the ultrasound?

during ultrasound examination of acute abdomen, sometimes we see free fluid , can we know the type of fluid ,whether blood or not , using ultrasound only? if yes , what are the sonographic...

01 March 2021 7,476 4 View

The reason that the culture of white fungi Contamination with green mold(Trichoderma), the culture autoclave at121 for 15 min. What is it?

Is the period to autoclave not enough? The inoculation in a hood and flame and UV and alcohol.

27 February 2021 9,356 3 View

Is there a fast method matching citations to bibliography (manually typed references)?

I need to be able to match in-text citations to a reference list and back again in very large documents (100+ pages) WITHOUT the use of referencing software like Endnote. my method is to type the...

27 February 2021 9,848 1 View

Can I put a figure that was drawn by Bio-Render in publication paper?

I want to publish paper in journal but I have figures which have drawn with bio-renders, can I put them in the paper or which other software I can use to draw them?

27 February 2021 8,179 1 View

What is the most suitable medium of ascomycetes fungi ??

26 February 2021 4,413 1 View

Is it possible to set a breakpoint at a specific DYNAMIC instruction using the GNU Debugger (GDB)?

Using GDB, it is straightforward to debug and monitor a target program by setting a break-point at a specific instruction, since the instruction addresses are known. However, we have two...

24 February 2021 1,598 2 View

Is it possible that Cancer cells could survive in heparinized blood for 48 hours?

Hi everyone, I am just wondering if we incubated 200 microliter of whole blood with 50 microliter of cancer cells(1bout 5000cells) do you think that cancer would survive for 48 hours?

23 February 2021 3,840 3 View

What is the most prevalent drug related problem in the developed countries?

Detecting the drug related problems is an important step in the pharmaceutical care plan

21 February 2021 5,421 3 View

What are the dimensions by which the independent variable (big data) can be measured, Knowing We using the questionnaire instrument?

We have a research project consisting of three variables where big data is the independent variable. Some previous studies showed that Volume, Variety, Velocity and Veracity Are the dimensions...

21 February 2021 8,133 2 View

Is nanotechnology part of materials science? Is it possible to obtain Pico technology and move from hypothesis to application?

Pico-scale This is three orders of magnitude smaller than a nanometer (and thus most nanotechnology) and two orders of magnitude smaller than most chemistry transformations and measurements. Pico...

18 February 2021 4,540 5 View

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance

03 March 2021 3,568 1 View

Gel electrophoresis result for total RNA samples, 1.5 Agarose gel was used, How to improve the integrity of RNA?

Gel electrophoresis, RNA degradation, RNA extraction from fresh tissue

02 March 2021 5,433 5 View

Is There Any Feasible Method To Test Fluorescent Compounds Other Than Fluorescence Spectrometers ??

Is There Any Feasible Method To Test The Efficiency Of Fluorescent Compounds Other Than UV Spectrometers ? Suggestions Would Be Appreciated !

02 March 2021 5,785 3 View

Ester cleavage conditions?

Hi all, I need to cleave the ester in Ethyl 2-amino-1H-imidazole-5-carboxylate to make the carboxylic acid and retrieve the product from solution to use in downstream applications of linking to...

01 March 2021 8,766 3 View

Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...

01 March 2021 8,544 1 View

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 3 View

Expression cloning by using cDNA,do I need to modify the cDNA first?

I am going to have a expression cloning of mammalian gene by using shuttle plasmid to transforming the E.coli However I don't know I should only inserting the Coding sequence ,or I can...

28 February 2021 5,440 3 View

Problem with shRNA transfection. Why are only two plates getting contaminated instead of all?

I transfected my LNCaP-WT cells with 3 shRNA plus their NTC two weeks ago and split two puromycin selected cell plates on Friday last week(Feb 26). I checked for GFP in the cells, and they all...

28 February 2021 4,949 3 View

Why am I not able to use all VEP programs for the in silico analysis of avpr1b SNP's?

Good day researchers I am busy analysing the predicted effects of certain SNP's in AVPR1b. When using the VEP prediction programs (SIFT, Polyphen, FATHMM, LRT, Provean, Mutation Assessor, and...

28 February 2021 5,197 1 View

What is the difference between genotype and haplotype data in the ped format?

Hi everybody In the ped format for genotype, alleles of any SNP are represented by two columns (one for each allele, separated by a space). Is a column sufficient for the haplotype to...

27 February 2021 1,965 1 View