Converting cDNA to Protein Sequence: Methods and Recommendations

[gravenewworld]

Has anyone here done this before? If so, kit recommendations and protocol? Any help would be awesome. Thanks!

 

[Ygggdrasil]

What exactly do you want to do? Do you have a cDNA sequence and you want to figure out the protein sequence that it codes for?

 

[gravenewworld]

Yes exactly. Getting the cDNA is easy, I have experience with that for PCR, but instead of doing PCR, I was wondering how you can use your cDNA to get the protein sequence. My PI said it should be easily doable. I'm not sure how though.

 

[Ygggdrasil]

Once you have the cDNA sequence, you just need to sequence it (send a sample of the cDNA along with the appropriate primers to a DNA sequencing facility/company). You can the just read the protein sequence from the sequence you get (for example, using an online translation tool such as http://www.bioinformatics.org/sms2/translate.html

Let me know if you want more detail on any of these steps.

 

[LudusRex]

Yes, I have experience with converting cDNA to protein sequence. The most common method is to use a commercial cDNA synthesis kit, such as the SuperScript III First-Strand Synthesis System. This kit includes all the necessary reagents and protocols for efficient cDNA synthesis. As for the protocol, it involves reverse transcription of RNA using a reverse transcriptase enzyme, followed by amplification of the cDNA using polymerase chain reaction (PCR). Once the cDNA is amplified, it can be sequenced and translated into a protein sequence using bioinformatics tools. I recommend consulting the manufacturer's instructions for specific details on the protocol. I hope this helps!