README File: 

Title: Sequence terminus dependent PCR for site-specific mutation and modification detection

Authors: Gaolian Xu, Hao Yang, Jiani Qiu,  Julien Reboud, Linqing Zhen, Wei Ren, Hong Xu, Jonathan M. Cooper, Hongchen Gu
 
This readme file aims to provide a guide to use the data associated with this publication, available here in the following folders.  

Note: Files with the extensions .ab1 are sequencing results and can be read with software Chromas (http://technelysium.com.au/wp/chromas/ v2.6.6 accessed 6th February 2022) or 4peaks (https://nucleobytes.com/4peaks/index.html v1.8, accessed 6th February 2022)

Figure 1: 
	'Figure 1.xlsx' contains the data associated with Figure 1d and Figure 1e as two separate sheets: 
	In Figure 1d, column 1 is the temperature, whilst columns 2-5 contain the first derivative of the fluorescence intensity for different lengths (stated in nt in the first row -3)
	In Figure 1e, the first column provides the cycle value. The rest of the columns provide the fluorescent signal for different conditions and replicates annotated with the concentration as follows: 100p Ð p stands for pM. When f is used, it stands for fM.  

Figure 2: 
'Figure 2.xlsx' contains the data associated with Figure 2b-d as separate sheets
      Figure 2b and 2c present the raw image of the gels with markers.
	Figure 2d contains the repeats of the data, as a function of the concentration of methylated DNA (%)
      'Figure 2e.ab1' contains the trace of the sequencing results presented in Figure 2e.

Figure 3: 
'Figure 3.xlsx' contains the data associated with Figure 3a-b as separate sheets
	In both sheets, the first column is the cycle number whilst other columns are fluorescence intensity of different template concentrations (in copies/reaction) Ð NC is negative control. 

Figure 4: 'Figure 4b.xlsx' presents the fluorescent intensity as function of cycle number (column 1) for different target concentration and samples. 'NEG' is negative.  

Figure 5: 
	'Figure 5.xlsx' contains the data associated with Figure 5b-c as separate sheets
	Figure 5b presents the raw data of fluorescence intensity for each sample. The first column is cycle number and samples are identified by name. Data starts at row 3 
	Figure 5c presents the raw image of the gel with markers.
      'Figure 5d.ab1' contains the trace of the sequencing results presented in Figure 5d.

Figure 6: 
	'Figure 6.xlsx' contains the data associated with Figure 6b-e as separate sheets
	Figure 6b presents the raw image of the gel with markers.
      Figure 6c presents the raw data of fluorescence intensity for each sample. The first column is cycle number and samples are identified by number of base pairs. Data starts at row 4.  The inset gel is presented with markers starting at column O. 
      Figure 6d and 6e present the fluorescence intensity as a function of cycle number (column 1). Different samples represent different template concentrations. 
	
Figure S1: 
      'Figure S1.xlsx' provides the raw data of deltaG analysis as a function of stem length (in bp). The first three rows present the denomination of the columns. Data starts at row 4. 

Figure S3: 
	'Figure S3a.xlsx' provides the raw data of fluorescence signal for different lengths and target concentration in each column (apart from first column is cycle number). Data is in triplicate for both parameters. Columns are identifiable with length and target concentration. For example column 2 is 'A2: STEM 15-100P', referring to length of 15bp and a concentration of 100pM. 
	'Figure S3b.xlsx' follows the same nomenclature. 

Figure S4:
	'Figure S4.xlsx' presents the raw image of the gel with markers.

Figure S5:
	'Figure S5a.ab1',  'Figure S5b.ab1' and 'Figure S5c.ab1' contain the trace of the sequencing results presented in Figure S5a-c. 

Figure S6:
      'Figure S6.xlsx' contains the data for Figure S6a in columns 2-4 and for Figure S6b in columns 8-11.

Figure S9
	'Figure S9.xlsx' presents the fluorescence intensity versus cycle number (column 2) for different concentration of target (%) in columns 3-7.

Figure S12: 
	'Figure S12.xlsx' contains the data associated with Figure S12a-b as two separate sheets with the same format, with cycle number in column 2 and fluorescence intensity for different shift in length or position in columns 3 onwards. The last column is a negative sample.

Figure S13: 
	'Figure S13.xlsx' presents the fluorescence intensity as a function of cycle number (column 1) for different conditions represented by either 'one site' or 'two sites' as the 2 different series, as well as the amount tested. For example 'H2: one site 5ng' is the data for 'one site' and an amount of 5ng.