Your picture is too small to see. But, in general practice, you can clone your PCR product into a vector such as TOPO cloning vector (see attached, they have a TA cloning site, and also have some restriction sites around), and use T7 or M13 sites on the vector to sequence your PCR product. You can sequence either from the T7 direction or M13 direction.

[update**: I think the earlier version of the question has no 'without cloning' words]

Hi Yuan

Sorry for that, please have a look at the attached file now. 

Hi Pranay,

1. Do you have a vector (with T7 and M13 sites) in mind or available which you want to clone in your PCR product?

2. What kind of DNA polymerase did you use for PCR amplification? If a regular Taq, you can even use a simple TA cloning vector to clone for sequencing.

3. If your PCR product is only for sequencing purpose, you can even perform direct sequencing without cloning

Dear Yuan

Could you please explain in detail how to do direct sequencing of PCR product with sticky ends. I have introduced 2 restriction sites HindIII at 5' end of forward PCR primer and BglII at 5' end of  reverse PCR to amplify a 303 bp fragment now I want to sequence the product with overhangs using sequencing primers T7 and M13 since my previous primers are not suitable for direct sequencing. 

1. In principle, for direct PCR product sequencing:

step 1: PCR,

step 2: purification,

step 3: use either one of the primer (which you used for the PCR amplification) for sequencing (if you are using BigDye for sequencing, there should be an instruction for you to set up sequencing reaction for sequencing). You can set up 2 reactions, one use the Forward primer, and the other one use the Reverse primer to sequence from both directions.

2. If you don't have T7 or M13 sites (sequences) on your PCR product, you cannot use T7 or M13 primer for direct sequencing.

3. Why the previous primers are not suitable for direct sequencing? 

4. You can re-design the sequencing primers using the sequences at the end of the PCR product

By attaching the adaptors with complementary overhangs made from T7 and M13 Reverse primers sequences we can make our PCR product compatible for Sequencing primers.

Kindly look at the attached picture

Thank you 

Yes, of course you can do that. First, add the adaptors and then you can use T7 or M13 primers for sequencing.

But, my point is: why bother going through this ligation of your PCR product and the adaptors (if you just want to check the sequence of your PCR product)? You can simply re-order two primers (see attached PPT: the green F1 and yellow R1), about 18 mers (your choose). Then, you are on business, doing your sequencing. You can use these primers for direct sequencing.

using pcr primers can be dangerous for direct sequencing if the targeted region is heterozygous like MHC. suppose u get a homozygous sequence then its ok, however many a time u may end up with heterozygou sequence. in that case after pcr direct sequencing you need to again go for cloning and sequencing and then deductional prediction of the heterozygous sequence.